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Sample records for bunyavirus

  1. Bunyaviruses

    Czech Academy of Sciences Publication Activity Database

    Kilian, Patrik; Danielová, V.; Růžek, Daniel

    Florida : CRC Press, 2013, s. 67-91. ISBN 9781466567207 Institutional support: RVO:60077344 Keywords : RNA viruses * retroviruses * Bunyaviruses Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine

  2. Early Bunyavirus-Host Cell Interactions

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    Amelina Albornoz

    2016-05-01

    Full Text Available The Bunyaviridae is the largest family of RNA viruses, with over 350 members worldwide. Several of these viruses cause severe diseases in livestock and humans. With an increasing number and frequency of outbreaks, bunyaviruses represent a growing threat to public health and agricultural productivity globally. Yet, the receptors, cellular factors and endocytic pathways used by these emerging pathogens to infect cells remain largely uncharacterized. The focus of this review is on the early steps of bunyavirus infection, from virus binding to penetration from endosomes. We address current knowledge and advances for members from each genus in the Bunyaviridae family regarding virus receptors, uptake, intracellular trafficking and fusion.

  3. Early Bunyavirus-Host Cell Interactions.

    Science.gov (United States)

    Albornoz, Amelina; Hoffmann, Anja B; Lozach, Pierre-Yves; Tischler, Nicole D

    2016-01-01

    The Bunyaviridae is the largest family of RNA viruses, with over 350 members worldwide. Several of these viruses cause severe diseases in livestock and humans. With an increasing number and frequency of outbreaks, bunyaviruses represent a growing threat to public health and agricultural productivity globally. Yet, the receptors, cellular factors and endocytic pathways used by these emerging pathogens to infect cells remain largely uncharacterized. The focus of this review is on the early steps of bunyavirus infection, from virus binding to penetration from endosomes. We address current knowledge and advances for members from each genus in the Bunyaviridae family regarding virus receptors, uptake, intracellular trafficking and fusion. PMID:27213430

  4. Bunyaviruses with segmented glycoprotein genes and methods for generating these viruses

    OpenAIRE

    Kortekaas, J.A.; Wichgers Schreur, P.J.; Oreshkova, N.D.; Moormann, R. J. M.

    2014-01-01

    Bunyaviruses with segmented glycoprotein precursor genes and methods for generating these viruses Abstract: The invention relates to a bunyavirus, in which separated (NSm)Gn and Gc coding regions are functionally present on two different genome segments, preferably a bunyavirus that comprises a total of at least 4 genome segments. The invention further relates to methods for producing said bunyavirus, and to a composition comprising said bunyavirus and a suitable excipient.

  5. Emerging and Reemeriging Human Bunyavirus Infections and Climate Change

    Science.gov (United States)

    Sutherland, Laura J.; Anyamba, Assaf; LaBeaud, A. Desiree

    2013-01-01

    The Bunyaviridae family includes a growing number of viruses that have contributed to the burden of emerging and reemerging infectious diseases around the globe. Many of these viruses cause severe clinical outcomes in human and animal populations, the results of which can be detrimental to public health and the economies of affected communities. The threat to endemic and non-native regions is particularly high, and national and international public health agencies are often on alert. Many of the bunyaviruses cause severe clinical disease including hemorrhage, organ failure, and death leading to their high-risk classification. Hantaviruses and Rift Valley fever virus (RVFV) (genus Phlebovirus) are National Institute of Allergy and Infectious Diseases Category A priority pathogens in the United States. Viral hemorrhagic fevers, a classification that includes many bunyaviruses, are immediately notifiable in the European Union. The emergence of new and reemerging bunyaviruses has resulted in numerous human and animal fatalities. Outbreaks of Rift Valley fever (RVF) in East Africa (1997/1998, 2006/2007), Sudan (2007), Southern Africa (2008-2010), Kenya (1997/1998, 2006/2007) (Anyamba et al., 2009, 2010; Breiman et al., 2010; Grobbelaar et al., 2011; Woods et al., 2002) and Saudi Arabia & Yemen (2000, 2010) (Food and Agriculture Organization, 2000; Hjelle and Glass, 2000; Madani et al., 2003) and the emergence of Sin Nombre virus (1993) (Hjelle and Glass, 2000) and most recently Schmallenberg virus (2011) (DEFRA, 2012) are prime examples of the devastating and worldwide toll bunyaviruses have on health and economies. Climate variability (precipitation and temperature in particular) greatly influence the ecological conditions that drive arboviral disease outbreaks across the globe. Several human and animal disease outbreaks have been influenced by changes in climate associated with the El Niño Southern Oscillation (ENSO) phenomenon including the bunyaviruses RVFV and Sin

  6. [Concomitant activity of 2 bunyaviruses in horses in Argentina].

    Science.gov (United States)

    Cámara, A; Contigiani, M S; Medeot, S I

    1990-01-01

    A serologic survey of horses for Kairi (KRI) and Cache Valley (CV), two related Bunyaviruses, was conducted simultaneously in Cordoba and Santa Fe provinces, Argentina, during late 1983 and 1984. The prevalence of neutralizing antibodies only for KRI was 13.3% and only for CV was 40.0%; but if the total positive sera for KRI and CV were taken into account, the prevalence reached 48.3 and 75.0%, respectively. The prevalence for CV was higher than for KRI in Cordoba (p less than 0.01), but both were similar in Santa Fe province. The demonstration of seroconversion in horses of the two zones for both viruses indicates that these viruses have a concomitant activity. The infection rates (number of infections per 100 horses-month) were very high in Cordoba (4.4 and 7.1 for KRI and CV) but also in Santa Fe (2.9 and 9.5 for the two viruses respectively), without significant difference in each province. Despite this high activity, no signs of illness or death imputed to these viruses were registered, in these areas during the period of observation. This apparent absence of associated equine disease may be the consequence of the low or null virus pathogenicity or the underrecognition or underreporting of the clinical cases. PMID:2126879

  7. Multilamellar Structures and Filament Bundles Are Found on the Cell Surface during Bunyavirus Egress

    OpenAIRE

    Sanz-Sánchez, Laura; Risco, Cristina

    2013-01-01

    Inside cells, viruses build specialized compartments for replication and morphogenesis. We observed that virus release associates with specific structures found on the surface of mammalian cells. Cultured adherent cells were infected with a bunyavirus and processed for oriented sectioning and transmission electron microscopy. Imaging of cell basal regions showed sophisticated multilamellar structures (MLS) and extracellular filament bundles with attached viruses. Correlative light and electro...

  8. Brus Laguna virus, a Gamboa bunyavirus from Aedeomyia squamipennis collected in Honduras.

    Science.gov (United States)

    Calisher, C H; Lazuick, J S; Sudia, W D

    1988-10-01

    A virus isolate from Aedeomyia squamipennis collected in Honduras in 1967 was identified as a member of the Gamboa serogroup (family Bunyaviridae, genus Bunyavirus). This is the ninth Gamboa serogroup virus and the eighth shown to be a distinct serotype. PMID:2903690

  9. Multilamellar structures and filament bundles are found on the cell surface during bunyavirus egress.

    Directory of Open Access Journals (Sweden)

    Laura Sanz-Sánchez

    Full Text Available Inside cells, viruses build specialized compartments for replication and morphogenesis. We observed that virus release associates with specific structures found on the surface of mammalian cells. Cultured adherent cells were infected with a bunyavirus and processed for oriented sectioning and transmission electron microscopy. Imaging of cell basal regions showed sophisticated multilamellar structures (MLS and extracellular filament bundles with attached viruses. Correlative light and electron microscopy confirmed that both MLS and filaments proliferated during the maximum egress of new viruses. MLS dimensions and structure were reminiscent of those reported for the nanostructures on gecko fingertips, which are responsible for the extraordinary attachment capacity of these lizards. As infected cells with MLS were more resistant to detachment than control cells, we propose an adhesive function for these structures, which would compensate for the loss of adherence during release of new virus progeny.

  10. Risk assessment of human infection with a novel bunyavirus in China

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    Tamano Matsui

    2012-11-01

    Full Text Available Objective: To assess the public health risk of human infection from a novel bunyavirus – severe fever with thrombocytopenia syndrome virus (SFTSV – in China.Methods: The likelihood of disease spread and the magnitude of public health impact were assessed to clarify overall risk. Literature about hazard, exposure and contextual factors associated with SFTSV infection was collected and reviewed. Information on SFTSV cases and the population in six provinces under surveillance was compared.Results: SFTSV is a member of the Phlebovirus genus of the Bunyaviridae family. A widely distributed tick species, Haemaphysalis longicornis, can act as the vector; thus the disease is likely to spread in China. Symptoms of SFTSV infection are nonspecific, but have led to multiorgan dysfunction in severe cases. High-risk populations include farmers and older females. Evidence of human-to-human transmission within family and hospital has been reported. The capacity for treatment and diagnosis of SFTSV are adequate in rural communities in China, and community awareness of the disease should be high. Discussion: There is a low to moderate public health risk related to SFTSV human infection in China. There is potential for an increase in the number of cases reported as awareness increases and when surveillance is expanded.

  11. The genome sequence of Lone Star virus, a highly divergent bunyavirus found in the Amblyomma americanum tick.

    Science.gov (United States)

    Swei, Andrea; Russell, Brandy J; Naccache, Samia N; Kabre, Beniwende; Veeraraghavan, Narayanan; Pilgard, Mark A; Johnson, Barbara J B; Chiu, Charles Y

    2013-01-01

    Viruses in the family Bunyaviridae infect a wide range of plant, insect, and animal hosts. Tick-borne bunyaviruses in the Phlebovirus genus, including Severe Fever with Thrombocytopenia Syndrome virus (SFTSV) in China, Heartland virus (HRTV) in the United States, and Bhanja virus in Eurasia and Africa have been associated with acute febrile illness in humans. Here we sought to characterize the growth characteristics and genome of Lone Star virus (LSV), an unclassified bunyavirus originally isolated from the lone star tick Amblyomma americanum. LSV was able to infect both human (HeLa) and monkey (Vero) cells. Cytopathic effects were seen within 72 h in both cell lines; vacuolization was observed in infected Vero, but not HeLa, cells. Viral culture supernatants were examined by unbiased deep sequencing and analysis using an in-house developed rapid computational pipeline for viral discovery, which definitively identified LSV as a phlebovirus. De novo assembly of the full genome revealed that LSV is highly divergent, sharing identity with any other bunyavirus. Despite this sequence diversity, LSV was found by phylogenetic analysis to be part of a well-supported clade that includes members of the Bhanja group viruses, which are most closely related to SFSTV/HRTV. The genome sequencing of LSV is a critical first step in developing diagnostic tools to determine the risk of arbovirus transmission by A. americanum, a tick of growing importance given its expanding geographic range and competence as a disease vector. This study also underscores the power of deep sequencing analysis in rapidly identifying and sequencing the genomes of viruses of potential clinical and public health significance. PMID:23637969

  12. Proteomics computational analyses suggest that the carboxyl terminal glycoproteins of Bunyaviruses are class II viral fusion protein (beta-penetrenes

    Directory of Open Access Journals (Sweden)

    Garry Robert F

    2004-11-01

    Full Text Available Abstract The Bunyaviridae family of enveloped RNA viruses includes five genuses, orthobunyaviruses, hantaviruses, phleboviruses, nairoviruses and tospoviruses. It has not been determined which Bunyavirus protein mediates virion:cell membrane fusion. Class II viral fusion proteins (beta-penetrenes, encoded by members of the Alphaviridae and Flaviviridae, are comprised of three antiparallel beta sheet domains with an internal fusion peptide located at the end of domain II. Proteomics computational analyses indicate that the carboxyl terminal glycoprotein (Gc encoded by Sandfly fever virus (SAN, a phlebovirus, has a significant amino acid sequence similarity with envelope protein 1 (E1, the class II fusion protein of Sindbis virus (SIN, an Alphavirus. Similar sequences and common structural/functional motifs, including domains with a high propensity to interface with bilayer membranes, are located collinearly in SAN Gc and SIN E1. Gc encoded by members of each Bunyavirus genus share several sequence and structural motifs. These results suggest that Gc of Bunyaviridae, and similar proteins of Tenuiviruses and a group of Caenorhabditis elegans retroviruses, are class II viral fusion proteins. Comparisons of divergent viral fusion proteins can reveal features essential for virion:cell fusion, and suggest drug and vaccine strategies.

  13. Molecular characterization of human pathogenic bunyaviruses of the Nyando and Bwamba/Pongola virus groups leads to the genetic identification of Mojui dos Campos and Kaeng Khoi virus.

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    Allison Groseth

    2014-09-01

    Full Text Available Human infection with Bwamba virus (BWAV and the closely related Pongola virus (PGAV, as well as Nyando virus (NDV, are important causes of febrile illness in Africa. However, despite seroprevalence studies that indicate high rates of infection in many countries, these viruses remain relatively unknown and unstudied. In addition, a number of unclassified bunyaviruses have been isolated over the years often with uncertain relationships to human disease.In order to better understand the genetic and evolutionary relationships among orthobunyaviruses associated with human disease, we have sequenced the complete genomes for all 3 segments of multiple strains of BWAV (n = 2, PGAV (n = 2 and NDV (n = 4, as well as the previously unclassified Mojuí dos Campos (MDCV and Kaeng Khoi viruses (KKV. Based on phylogenetic analysis, we show that these viruses populate 2 distinct branches, one made up of BWAV and PGAV and the other composed of NDV, MDCV and KKV. Interestingly, the NDV strains analyzed form two distinct clades which differed by >10% on the amino acid level across all protein products. In addition, the assignment of two bat-associated bunyaviruses into the NDV group, which is clearly associated with mosquito-borne infection, led us to analyze the ability of these different viruses to grow in bat (RE05 and Tb 1 Lu and mosquito (C6/36 cell lines, and indeed all the viruses tested were capable of efficient growth in these cell types.On the basis of our analyses, it is proposed to reclassify the NDV strains ERET147 and YM176-66 as a new virus species. Further, our analysis definitively identifies the previously unclassified bunyaviruses MDCV and KKV as distinct species within the NDV group and suggests that these viruses may have a broader host range than is currently appreciated.

  14. Caracterização e relacionamento antigênico de três novos Bunyavirus no grupo Anopheles A (Bunyaviridae dos arbovirus Characterization and antigenic relationship of three new Bunyavirus in the Anopheles A serogroup (Bunyaviridae of arboviruses

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    Jorge Fernando Soares Travassos da Rosa

    1992-06-01

    Full Text Available São descritos o isolamento e a caracterização de três novos arbovirus isolados na região da Usina Hidro-Elétrica de Tucuruí (UHE-TUC. Os três novos arbovirus pertencem ao grupo Anopheles A(ANA, gênero Bunyavirus (família Bunyaviridae. Os vírus Tucuruí (TUC, Caraipé (CPE e Arumateua (ART são relacionados entre si e com o vírus Trombetas (TBT, formando dentro do grupo ANA um complexo chamado Trombetas. Os arbovirus TUC, CPE e ART foram obtidos a partir de lotes de mosquitos Anopheles (Nyssorhynchus sp capturados em Tucuruí, nas proximidades da usina hidrelétrica de Tucuruí, Estado do Pará, nos meses de fevereiro, agosto e outubro de 1984, respectivamente. Até o final de 1990 os vírus TUC, CPE e ART foram isolados 12, 32 e 28 vezes respectivamente, sempre na região da UHE-TUC, exceção feita ao vírus TUC, do qual se obteve uma amostra procedente de Balbina, onde também foi construída uma hidroelétrica. Até o presente, esses vírus só foram isolados a partir de mosquitos do grupo An. (Nys. principalmente, a partir das espécies An. (Nys. nuneztovari e An. (Nys. triannulatus também consideradas vetores secundários da malária na Amazônia Brasileira. Testes sorológicos executados com soros humanos e de diversas espécies de animais silvestres foram negativos, com exceção de um soro de um carnívoro de espécie Nasua nasua que neutralizou a amostra TUC em títulos de 2.6 índice logaritmico de neutralização (ILN.The isolation and characterization of three new viruses obtained from the Tucuruí hydroelectric dam region is repeated. These three agents belong to the Anopheles A serogroup, genus Bunyavirus, Bunyaviridae. The Tucuruí (TUC, Caraipe (CPE and Arumateua (ART viruses have close relationships with each other and with Trombetas (TBT virus, an Anopheles A virus previously isolated in the Amazon Region of Brazil. These viruses form the "Trombetas complex". TUC, CPE and ART viruses were obtained from pools of

  15. Ultrastructural, Antigenic and Physicochemical Characterization of the Mojuí dos Campos (Bunyavirus Isolated from Bat in the Brazilian Amazon Region

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    Wanzeller Ana LM

    2002-01-01

    Full Text Available The Mojuí dos Campos virus (MDCV was isolated from the blood of an unidentified bat (Chiroptera captured in Mojuí dos Campos, Santarém, State of Pará, Brazil, in 1975 and considerated to be antigenically different from other 102 arboviruses belonging to several antigenic groups isolated in the Amazon region or another region by complement fixation tests. The objective of this work was to develop a morphologic, an antigenic and physicochemical characterization of this virus. MDCV produces cytopathic effect in Vero cells, 24 h post-infection (p.i, and the degree of cellular destruction increases after a few hours. Negative staining electron microscopy of the supernatant of Vero cell cultures showed the presence of coated viral particles with a diameter of around 98 nm. Ultrathin sections of Vero cells, and brain and liver of newborn mice infected with MDCV showed an assembly of the viral particles into the Golgi vesicles. The synthesis kinetics of the proteins for MDCV were similar to that observed for other bunyaviruses, and viral proteins could be detected as early as 6 h p.i. Our results reinforce the original studies which had classified MDCV in the family Bunyaviridae, genus Bunyavirus as an ungrouped virus, and it may represent the prototype of a new serogroup.

  16. Caracterização e relacionamento antigênico de três novos Bunyavirus no grupo Anopheles A (Bunyaviridae) dos arbovirus

    OpenAIRE

    Rosa Jorge Fernando Soares Travassos da; Rosa Amélia Paes de Andrade Travassos da; Dégallier Nicolas; Vasconcelos Pedro Fernando da Costa

    1992-01-01

    São descritos o isolamento e a caracterização de três novos arbovirus isolados na região da Usina Hidro-Elétrica de Tucuruí (UHE-TUC). Os três novos arbovirus pertencem ao grupo Anopheles A(ANA), gênero Bunyavirus (família Bunyaviridae). Os vírus Tucuruí (TUC), Caraipé (CPE) e Arumateua (ART) são relacionados entre si e com o vírus Trombetas (TBT), formando dentro do grupo ANA um complexo chamado Trombetas. Os arbovirus TUC, CPE e ART foram obtidos a partir de lotes de mosquitos Anopheles (Ny...

  17. Molecular Characterization of Human Pathogenic Bunyaviruses of the Nyando and Bwamba/Pongola Virus Groups Leads to the Genetic Identification of Mojuí dos Campos and Kaeng Khoi Virus

    Science.gov (United States)

    Groseth, Allison; Mampilli, Veena; Weisend, Carla; Dahlstrom, Eric; Porcella, Stephen F.; Russell, Brandy J.; Tesh, Robert B.; Ebihara, Hideki

    2014-01-01

    Background Human infection with Bwamba virus (BWAV) and the closely related Pongola virus (PGAV), as well as Nyando virus (NDV), are important causes of febrile illness in Africa. However, despite seroprevalence studies that indicate high rates of infection in many countries, these viruses remain relatively unknown and unstudied. In addition, a number of unclassified bunyaviruses have been isolated over the years often with uncertain relationships to human disease. Methodology/Principal Findings In order to better understand the genetic and evolutionary relationships among orthobunyaviruses associated with human disease, we have sequenced the complete genomes for all 3 segments of multiple strains of BWAV (n = 2), PGAV (n = 2) and NDV (n = 4), as well as the previously unclassified Mojuí dos Campos (MDCV) and Kaeng Khoi viruses (KKV). Based on phylogenetic analysis, we show that these viruses populate 2 distinct branches, one made up of BWAV and PGAV and the other composed of NDV, MDCV and KKV. Interestingly, the NDV strains analyzed form two distinct clades which differed by >10% on the amino acid level across all protein products. In addition, the assignment of two bat-associated bunyaviruses into the NDV group, which is clearly associated with mosquito-borne infection, led us to analyze the ability of these different viruses to grow in bat (RE05 and Tb 1 Lu) and mosquito (C6/36) cell lines, and indeed all the viruses tested were capable of efficient growth in these cell types. Conclusions/Significance On the basis of our analyses, it is proposed to reclassify the NDV strains ERET147 and YM176-66 as a new virus species. Further, our analysis definitively identifies the previously unclassified bunyaviruses MDCV and KKV as distinct species within the NDV group and suggests that these viruses may have a broader host range than is currently appreciated. PMID:25188437

  18. Caracterização e relacionamento antigênico de três novos Bunyavirus no grupo Anopheles A (Bunyaviridae dos arbovirus

    Directory of Open Access Journals (Sweden)

    Rosa Jorge Fernando Soares Travassos da

    1992-01-01

    Full Text Available São descritos o isolamento e a caracterização de três novos arbovirus isolados na região da Usina Hidro-Elétrica de Tucuruí (UHE-TUC. Os três novos arbovirus pertencem ao grupo Anopheles A(ANA, gênero Bunyavirus (família Bunyaviridae. Os vírus Tucuruí (TUC, Caraipé (CPE e Arumateua (ART são relacionados entre si e com o vírus Trombetas (TBT, formando dentro do grupo ANA um complexo chamado Trombetas. Os arbovirus TUC, CPE e ART foram obtidos a partir de lotes de mosquitos Anopheles (Nyssorhynchus sp capturados em Tucuruí, nas proximidades da usina hidrelétrica de Tucuruí, Estado do Pará, nos meses de fevereiro, agosto e outubro de 1984, respectivamente. Até o final de 1990 os vírus TUC, CPE e ART foram isolados 12, 32 e 28 vezes respectivamente, sempre na região da UHE-TUC, exceção feita ao vírus TUC, do qual se obteve uma amostra procedente de Balbina, onde também foi construída uma hidroelétrica. Até o presente, esses vírus só foram isolados a partir de mosquitos do grupo An. (Nys. principalmente, a partir das espécies An. (Nys. nuneztovari e An. (Nys. triannulatus também consideradas vetores secundários da malária na Amazônia Brasileira. Testes sorológicos executados com soros humanos e de diversas espécies de animais silvestres foram negativos, com exceção de um soro de um carnívoro de espécie Nasua nasua que neutralizou a amostra TUC em títulos de 2.6 índice logaritmico de neutralização (ILN.

  19. Caracterização e relacionamento antigênico de três novos Bunyavirus no grupo Anopheles A (Bunyaviridae dos arbovirus

    Directory of Open Access Journals (Sweden)

    Jorge Fernando Soares Travassos da Rosa

    1992-06-01

    Full Text Available São descritos o isolamento e a caracterização de três novos arbovirus isolados na região da Usina Hidro-Elétrica de Tucuruí (UHE-TUC. Os três novos arbovirus pertencem ao grupo Anopheles A(ANA, gênero Bunyavirus (família Bunyaviridae. Os vírus Tucuruí (TUC, Caraipé (CPE e Arumateua (ART são relacionados entre si e com o vírus Trombetas (TBT, formando dentro do grupo ANA um complexo chamado Trombetas. Os arbovirus TUC, CPE e ART foram obtidos a partir de lotes de mosquitos Anopheles (Nyssorhynchus sp capturados em Tucuruí, nas proximidades da usina hidrelétrica de Tucuruí, Estado do Pará, nos meses de fevereiro, agosto e outubro de 1984, respectivamente. Até o final de 1990 os vírus TUC, CPE e ART foram isolados 12, 32 e 28 vezes respectivamente, sempre na região da UHE-TUC, exceção feita ao vírus TUC, do qual se obteve uma amostra procedente de Balbina, onde também foi construída uma hidroelétrica. Até o presente, esses vírus só foram isolados a partir de mosquitos do grupo An. (Nys. principalmente, a partir das espécies An. (Nys. nuneztovari e An. (Nys. triannulatus também consideradas vetores secundários da malária na Amazônia Brasileira. Testes sorológicos executados com soros humanos e de diversas espécies de animais silvestres foram negativos, com exceção de um soro de um carnívoro de espécie Nasua nasua que neutralizou a amostra TUC em títulos de 2.6 índice logaritmico de neutralização (ILN.

  20. Establishment of the culture process for scale production of SFTS Bunyavirus%发热伴血小板减少综合征布尼亚病毒规模化生产培养工艺的建立

    Institute of Scientific and Technical Information of China (English)

    杨骏宇; 姜东林; 陈小芳; 承静; 刘培生; 金坚

    2013-01-01

    Objective To investigate the ideal culture process of SFTS Bunyavirus and provide the basic date for scale pro-duction.Methods Vero cells were cultured using cell factory .The working seed of SFTS bunyavirus was inoculated into Vero cells when the cells grew into monolayer .The virus were harvested with continuous perfused culture system or harves-ting the supernatant of culture when the cells achieved fully lesions .The virus titers and antigen contents were used as indi-cators for selecting medium , pH of medium , added human serum albumin concentration ,virus incubation temperature ,cells age and MOI of virus.Results 3-4 days old Vero cells were inoculated with 0.01-0.001,MOI of SFTS Bunyavirus, DMEM solution containing 0.3%human serum albumin was selected as the medium (pH 7.6-7.8), the virus were cul-tured at 35 ℃for 7 days, the harvested virus titer is 7.87 LgCCID50/mL and the antigen content is 170.1 μg/mL.Con-clusion The scale culture process of SFTS Bunyavirus was initially established , and it laid a good foundation for industrial production .%目的:通过对发热伴血小板减少综合征布尼亚病毒(简称“新布尼亚病毒”)进行规模化生产培养工艺研究,为新布尼亚病毒规模化生产提供有力支持。方法采用细胞工厂培养Vero细胞,待其长成致密单层后,取工作种子批新布尼亚病毒毒种接种细胞,采用连续收获或细胞病变充分时收获培养液上清的方法收获病毒,并以病毒滴度、抗原含量作为评价指标选择基础培养基、培养基pH、人血白蛋白添加浓度、接种细胞日龄、接种病毒MOI以及病毒培养温度。结果按0.01~0.001 MOI接种3~4日龄Vero细胞,病毒培养液选择含0.3%人血白蛋白pH 7.6~7.8的DMEM溶液,35℃培养7 d收获,病毒收获液病毒滴度7.87 LgCCID50/mL、抗原含量170.1μg/mL。结论初步建立了新布尼亚病毒规模化生产培养工艺,为后续工业化生产提供了数据支持。

  1. Chapter 30. Other Bunyavirus Infections

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    Rift Valley fever virus (RVFV) is a mosquito-transmitted virus or arbovirus that is endemic in sub-Saharan Africa. In the last decade, Rift Valley fever (RVF) outbreaks have resulted in loss of human and animal life, as well as had significant economic impact. The disease in livestock is primarily a...

  2. TaqMan探针实时荧光定量反转录-聚合酶链反应检测新布尼亚病毒L基因方法的建立与应用%Establishment of real time RT-PCR assay for detecting novel Bunyavirus L gene

    Institute of Scientific and Technical Information of China (English)

    虞吉寅; 王忠发; 任宜; 毛彬

    2013-01-01

    Objective To develop a sensitive and specific real time reverse transcription PCR ( RT-PCR) assay for nucleic acid detection of novel Bunyavirus. Methods Primers and TaqMan probe were designed according to the L gene of novel Bunyavirus and the real time RT-PCR was used to detect novel Bunyavirus from serum and plasma samples of 50 clinical suspected patients and from serum of 25 healthy people. Two virus strains isolated from the patients' serum were used to evaluate the specificity and sensitivity as well as the reproducibility of the established real time RT-PCR assay. Results The established real time RT-PCR assay had high specificity, with the Ct values showing good linear relation to the copy numbers of template DNA in the standard curve (R2 =0.999). In this assay, the target gene could be detected if the template DNA is more than 25 copies in 25 ΜL PCR reaction, indicating that the sensitive detection limit was 1 copy/μl. The reproducibility tests indicated that the intraassay coefficient of variation were all lower than 1. 0% . Among the 50 patients' serum samples, 30 were positive, 25 control' s serum samples and 28 rats lung and testes samples were all negative. Two of 8 blood samples were positive in virus culture. Conclusion The established real time RT-PCR assay was with high specificity, sensitivity and reproducibility, which could be used in the rapid nucleic acid detection of novel Bunyavirus in humans and animals and applied in the preliminary identification of the virus after isolation and culture.%目的 建立敏感、特异、实时荧光定量反转录-聚合酶链反应(real time fluorescent quantitative reverse transcription-polymerase chain reaction,rRT-PCR)方法,用于新布尼亚病毒的核酸检测.方法 以新布尼亚病毒L基因为靶基因设计引物以及TaqMan探针,建立real-time RT-PCR方法,并对舟山市岱山县50份临床疑似患者血清、血浆、25份正常人群血清和从患者血清中分离到的2株病毒

  3. Development of an improved reverse genetics system for Akabane bunyavirus.

    Science.gov (United States)

    Takenaka-Uema, Akiko; Sugiura, Keita; Bangphoomi, Norasuthi; Shioda, Chieko; Uchida, Kazuyuki; Kato, Kentaro; Haga, Takeshi; Murakami, Shin; Akashi, Hiroomi; Horimoto, Taisuke

    2016-06-01

    Akabane disease, caused by the insect-transmitted Akabane virus (AKAV), affects livestock by causing life-threatening deformities or mortality of fetuses. Therefore, Akabane disease has led to notable economic losses in numerous countries, including Japan. In this short communication, a new T7 RNA polymerase-based AKAV reverse genetics system was developed. Using this system, in which three plasmids transcribing antigenomic RNAs were transfected into cells stably expressing T7 polymerase, we successfully reconstituted the live attenuated vaccine TS-C2 strain (named rTTT), and also generated a mutant AKAV (rTTTΔNSs) that lacked the gene encoding the nonstructural NSs protein, which is regarded as a virulence factor. Analysis of growth kinetics revealed that rTTTΔNSs grew at a much slower rate than the rTTT and TS-C2 virus. These results suggest that our established reverse genetics system is a powerful tool that can be used for AKAV vaccine studies with gene-manipulated viruses. PMID:26927704

  4. Insights into bunyavirus architecture from electron cryotomography of Uukuniemi virus

    OpenAIRE

    Överby, A. K.; Pettersson, R F; Grünewald, K; Huiskonen, J.T.

    2008-01-01

    Bunyaviridae is a large family of viruses that have gained attention as “emerging viruses” because many members cause serious disease in humans, with an increasing number of outbreaks. These negative-strand RNA viruses possess a membrane envelope covered by glycoproteins. The virions are pleiomorphic and thus have not been amenable to structural characterization using common techniques that involve averaging of electron microscopic images. Here, we determined the three-dimensional structure o...

  5. Segment-specific terminal sequences of Bunyamwera bunyavirus regulate genome replication

    International Nuclear Information System (INIS)

    Bunyamwera virus (BUNV) is the prototype of both the Orthobunyavirus genus and the Bunyaviridae family of segmented negative sense RNA viruses. The tripartite BUNV genome consists of small (S), medium (M), and large (L) segments that are transcribed to give a single mRNA and replicated to generate an antigenome that is the template for synthesis of further genomic RNA strands. We modified an existing cDNA-derived RNA synthesis system to allow identification of BUNV RNA replication and transcription products by direct metabolic labeling. Direct RNA analysis allowed us to distinguish between template activities that affected either RNA replication or mRNA transcription, an ability that was not possible using previous reporter gene expression assays. We generated genome analogs containing the entire nontranslated terminal sequences of the S, M, and L BUNV segments surrounding a common sequence. Analysis of RNAs synthesized from these templates revealed that the relative abilities of BUNV segments to perform RNA replication was M > L > S. Exchange of segment-specific terminal nucleotides identified a 12-nt region located within both the 3' and 5' termini of the M segment that correlated with its high replication ability

  6. Synergistic antiviral effects of ribavirin and the C-nucleoside analogs tiazofurin and selenazofurin against togaviruses, bunyaviruses, and arenaviruses.

    OpenAIRE

    Huggins, J W; Robins, R K; Canonico, P G

    1984-01-01

    Binary combinations of the N-nucleoside ribavirin (1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide) and the C-nucleoside analog selenazofurin (2-beta-D-ribofuranosylselenazole-4-carboxamide) or tiazofurin (2-beta-D-ribofuranosylthiazole-4-carboxamide) were tested in vitro for activity against Venezuelan equine encephalomyelitis, Japanese encephalitis, yellow fever, Rift Valley fever, Korean hemorrhagic fever, and Pichinde viruses. The 50% effective dose for each compound alone or in a ser...

  7. AcEST: BP921688 [AcEST

    Lifescience Database Archive (English)

    Full Text Available MK0977 ... 31 2.6 sp|P04875|VGLM_BUNSH Envelope glycoprotein OS=Bunyavirus snowsho... 30 5.6 sp|A1U7G1|AMIE_...BUNSH Envelope glycoprotein OS=Bunyavirus snowshoe hare GN=M PE=1 SV=1 Length = 1441 Score = 29.6 bits (65),

  8. Nucleotide variability of Ťahyňa virus (Bunyaviridae, Orthobunyavirus) small (S) and medium (M) genomic segments in field strains differing in biological properties

    Czech Academy of Sciences Publication Activity Database

    Kilian, P.; Růžek, Daniel; Danielová, V.; Hypša, Václav; Grubhoffer, Libor

    2010-01-01

    Roč. 149, č. 1 (2010), s. 119-123. ISSN 0168-1702 R&D Projects: GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z60220518 Keywords : Tahyna virus * Bunyavirus * california group * genetic variability * virulence Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.905, year: 2010

  9. Evolutionary and phenotypic analysis of live virus isolates suggests arthropod origin of a pathogenic RNA virus family

    OpenAIRE

    Marklewitz, Marco; Zirkel, Florian; Kurth, Andreas; Drosten, Christian; Junglen, Sandra

    2015-01-01

    Knowledge of the origin and evolution of viruses provides important insight into virus emergence involving the acquisition of genes necessary for the infection of new host species or the development of pathogenicity. The family Bunyaviridae contains important arthropod-borne pathogens of humans, animals, and plants. In this study, we provide a comprehensive characterization of two novel lineages of insect-specific bunyaviruses that are in basal phylogenetic relationship to the rodent-borne ha...

  10. Lack of serologic evidence for an association between Cache Valley Virus infection and anencephaly and other neural tube defects in Texas.

    OpenAIRE

    Edwards, J. F.; Hendricks, K

    1997-01-01

    We tested the hypothesis that Cache Valley Virus (CVV), an endemic North American bunyavirus, may be involved in the pathogenesis of human neural tube defects. This investigation followed a 1990 and 1991 south Texas outbreak of neural tube defects with a high prevalence of anencephaly and the demonstration in 1987 that in utero infection by CVV was the cause of outbreaks of central nervous system and musculoskeletal defects in North American ruminants. Sera from 74 women who gave birth to inf...

  11. Oropuche virus: A virus present but ignored

    Directory of Open Access Journals (Sweden)

    Salim Mattar V.

    2015-09-01

    Full Text Available Bunyaviruses are RNA viruses that affect animals and plants; they have five genera and four of them affect humans: Orthobunyavirus, Nairovirus, Phlebovirus and Hantavirus. All of them are Arbovirus, except Hantavirus. The Orthobunyaviruses comprise Oropouche, Tahyna, La Crosse virus, California encephalitis virus and Heartland virus recently discovered (1. Except for Heartland virus which is transmitted by ticks of the genus Amblyoma, these Phleboviruses have as vectors mosquitoes, which bite small mammals which are able to be as reservoirs amplifiers.

  12. Transmission of La Crosse Virus in Southwest Virginia: Role of Accessory Vectors, Microfilarial Coinfection and Canine Seroprevalence

    OpenAIRE

    Troyano, Nancy Michelle

    2009-01-01

    Southwest Virginia has recently become an emerging focus of activity for La Crosse (LAC) virus, a mosquito-transmitted arbovirus in the California serogroup of Bunyaviruses. In 2005 and 2006, ovitrap surveys were conducted to access the spatiotemporal oviposition activity of LAC virus vectors Aedes triseriatus, Ae. albopictus and Ae. japonicus across a wide region of southwest Virginia. Egg abundance and oviposition patterns of these vectors were significantly different across the three stu...

  13. Forest disturbance, mosquito vector ecology and La Crosse virus dynamics in southwestern Virginia

    OpenAIRE

    Harris, Maria-Richetta Camille

    2014-01-01

    The influence of forest canopy disturbance (FCD) on La Crosse virus (LACV), leading cause of US pediatric arboviral encephalitis, is critical to understand in landscapes where forests are periodically harvested. Southwestern Virginia is part of an emerging focus of this interior forest bunyavirus. I investigated how the temperate forest mosquito community, LACV vectors, and the LACV amplifying vertebrate host (chipmunks) were impacted by logging. This research was conducted across an exper...

  14. Development of FGI-106 as a broad-spectrum therapeutic with activity against members of the family Bunyaviridae

    Directory of Open Access Journals (Sweden)

    Darci R Smith

    2010-02-01

    Full Text Available Darci R Smith1, Monica Ogg1, Aura Garrison1, Abdul Yunus2, Anna Honko1, Josh Johnson1, Gene Olinger1, Lisa E Hensley1, Michael S Kinch1United States Army Medical Research Institute of Infectious Diseases (USAMRII D, Fort Detrick, MD, USA; 2Functional Genetics, Inc., Gaithersburg, MD, USAAbstract: The family Bunyaviridae is a diverse group of negative-strand RNA viruses that infect a wide range of arthropod vectors and animal hosts. Based on the continuing need for new therapeutics to treat bunyavirus infections, we evaluated the potential efficacy of FGI-106, a small-molecular compound that previously demonstrated activity against different RNA viruses. FGI-106 displayed substantial antiviral activity in cell-based assays of different bunyavirus family members, including Asian and South American hantaviruses (Hantaan virus and Andes virus, Crimean-Congo hemorrhagic fever virus, La Crosse virus, and Rift Valley fever virus. The pharmacokinetic profile of FGI-106 revealed sufficient exposure of the drug to critical target organs (lung, liver, kidney, and spleen, which are frequently the sites of bunyavirus replication. Consistent with these findings, FGI-106 treatment delivered via intraperitoneal injection prior to virus exposure was sufficient to delay the onset of Rift Valley fever virus infection in mouse-based models and to enhance survival in the face of an otherwise lethal infection. Altogether, these results suggest a potential opportunity for the use of FGI-106 to treat infections by members of the family Bunyaviridae.Keywords: Rift Valley fever virus, bunyavirus, hantavirus, antiviral, therapeutic

  15. Serological Screening Suggests Presence of Schmallenberg Virus in Cattle, Sheep and Goat in the Zambezia Province, Mozambique

    OpenAIRE

    Blomström, A-L; Stenberg, H; Scharin, I; Figueiredo, J; Nhambirre, O; Abilio, A P; Fafetine, J; Berg, M.

    2014-01-01

    Schmallenberg virus (SBV) is a novel Orthobunyavirus within the family Bunyaviridae belonging to the Simbu serogroup. Schmallenberg virus infects ruminants and has since its discovery in the autumn 2011 been detected/spread to large parts of Europe. Most bunyaviruses are arboviruses, and SBV has been detected in biting midges in different European countries, suggesting that they may play a role in the transmission of the virus. It is not known how SBV was introduced to Europe and if SBV is pr...

  16. Transovarial transmission of Gamboa virus in a tropical mosquito, Aedeomyia squamipennis.

    Science.gov (United States)

    Dutary, B E; Petersen, J L; Peralta, P H; Tesh, R B

    1989-01-01

    We report transovarial transmission of Gamboa virus (Bunyavirus) in Aedeomyia squamipennis, a tropical mosquito which is active and bloodfeeding throughout the year. Gamboa virus was isolated during each of the 28 months of the study from every mosquito stage, including eggs, demonstrating that vertical transmission is a maintenance mechanism of this virus. The overall minimum infection rate was 5.1/1,000 mosquitoes. Identification of the 567 isolates by neutralization indicated that greater than or equal to 2 serotypes or subtypes of Gamboa virus circulate at the study site. PMID:2563641

  17. [Importation of rare but life-threatening and highly contagious diseases. Current situation and outlook].

    Science.gov (United States)

    Burchard, G D

    2015-10-01

    Internists should expect to be the first contact for patients with rare, but highly contagious, life-threatening illnesses. Although certainly not encountered often, it is associated with significant consequences. Thus, physicians should be familiar with viral hemorrhagic fevers: filoviruses cause Ebola and Marburg fever, arenaviruses cause Lassa fever and South American hemorrhagic fevers, and the bunyaviruses cause among others Crimean-Congo hemorrhagic fever. Furthermore, physicians should be familiar with highly contagious respiratory infections, such as hantavirus pulmonary syndrome, pneumonic plague, and Middle East respiratory syndrome (MERS). PMID:26391557

  18. Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF virus glycoproteins

    Directory of Open Access Journals (Sweden)

    Fernando Lisa

    2005-04-01

    Full Text Available Abstract Background Crimean-Congo Hemorrhagic Fever virus (CCHFV, a member of the genus Nairovirus, family Bunyaviridae, is a tick-borne pathogen causing severe disease in humans. To better understand the CCHFV life cycle and explore potential intervention strategies, we studied the biosynthesis and intracellular targeting of the glycoproteins, which are encoded by the M genome segment. Results Following determination of the complete genome sequence of the CCHFV reference strain IbAr10200, we generated expression plasmids for the individual expression of the glycoproteins GN and GC, using CMV- and chicken β-actin-driven promoters. The cellular localization of recombinantly expressed CCHFV glycoproteins was compared to authentic glycoproteins expressed during virus infection using indirect immunofluorescence assays, subcellular fractionation/western blot assays and confocal microscopy. To further elucidate potential intracellular targeting/retention signals of the two glycoproteins, GFP-fusion proteins containing different parts of the CCHFV glycoprotein were analyzed for their intracellular targeting. The N-terminal glycoprotein GN localized to the Golgi complex, a process mediated by retention/targeting signal(s in the cytoplasmic domain and ectodomain of this protein. In contrast, the C-terminal glycoprotein GC remained in the endoplasmic reticulum but could be rescued into the Golgi complex by co-expression of GN. Conclusion The data are consistent with the intracellular targeting of most bunyavirus glycoproteins and support the general model for assembly and budding of bunyavirus particles in the Golgi compartment.

  19. The Role of Phlebovirus Glycoproteins in Viral Entry, Assembly and Release.

    Science.gov (United States)

    Spiegel, Martin; Plegge, Teresa; Pöhlmann, Stefan

    2016-01-01

    Bunyaviruses are enveloped viruses with a tripartite RNA genome that can pose a serious threat to animal and human health. Members of the Phlebovirus genus of the family Bunyaviridae are transmitted by mosquitos and ticks to humans and include highly pathogenic agents like Rift Valley fever virus (RVFV) and severe fever with thrombocytopenia syndrome virus (SFTSV) as well as viruses that do not cause disease in humans, like Uukuniemi virus (UUKV). Phleboviruses and other bunyaviruses use their envelope proteins, Gn and Gc, for entry into target cells and for assembly of progeny particles in infected cells. Thus, binding of Gn and Gc to cell surface factors promotes viral attachment and uptake into cells and exposure to endosomal low pH induces Gc-driven fusion of the viral and the vesicle membranes. Moreover, Gn and Gc facilitate virion incorporation of the viral genome via their intracellular domains and Gn and Gc interactions allow the formation of a highly ordered glycoprotein lattice on the virion surface. Studies conducted in the last decade provided important insights into the configuration of phlebovirus Gn and Gc proteins in the viral membrane, the cellular factors used by phleboviruses for entry and the mechanisms employed by phlebovirus Gc proteins for membrane fusion. Here, we will review our knowledge on the glycoprotein biogenesis and the role of Gn and Gc proteins in the phlebovirus replication cycle. PMID:27455305

  20. The Role of Phlebovirus Glycoproteins in Viral Entry, Assembly and Release

    Directory of Open Access Journals (Sweden)

    Martin Spiegel

    2016-07-01

    Full Text Available Bunyaviruses are enveloped viruses with a tripartite RNA genome that can pose a serious threat to animal and human health. Members of the Phlebovirus genus of the family Bunyaviridae are transmitted by mosquitos and ticks to humans and include highly pathogenic agents like Rift Valley fever virus (RVFV and severe fever with thrombocytopenia syndrome virus (SFTSV as well as viruses that do not cause disease in humans, like Uukuniemi virus (UUKV. Phleboviruses and other bunyaviruses use their envelope proteins, Gn and Gc, for entry into target cells and for assembly of progeny particles in infected cells. Thus, binding of Gn and Gc to cell surface factors promotes viral attachment and uptake into cells and exposure to endosomal low pH induces Gc-driven fusion of the viral and the vesicle membranes. Moreover, Gn and Gc facilitate virion incorporation of the viral genome via their intracellular domains and Gn and Gc interactions allow the formation of a highly ordered glycoprotein lattice on the virion surface. Studies conducted in the last decade provided important insights into the configuration of phlebovirus Gn and Gc proteins in the viral membrane, the cellular factors used by phleboviruses for entry and the mechanisms employed by phlebovirus Gc proteins for membrane fusion. Here, we will review our knowledge on the glycoprotein biogenesis and the role of Gn and Gc proteins in the phlebovirus replication cycle.

  1. Thrips transmission of tospoviruses.

    Science.gov (United States)

    Rotenberg, Dorith; Jacobson, Alana L; Schneweis, Derek J; Whitfield, Anna E

    2015-12-01

    One hundred years ago, the disease tomato spotted wilt was first described in Australia. Since that time, knowledge of this disease caused by Tomato spotted wilt virus (TSWV) and transmitted by thrips (insects in the order Thysanoptera) has revealed a complex relationship between the virus, vector, plant host, and environment. Numerous tospoviruses and thrips vectors have been described, revealing diversity in plant host range and geographical distributions. Advances in characterization of the tripartite interaction between the virus, vector, and plant host have provided insight into molecular and ecological relationships. Comparison to animal-infecting viruses in the family Bunyaviridae has enabled the identification of commonalities between tospoviruses and other bunyaviruses in transmission by arthropod vectors and molecular interactions with hosts. This review provides a special emphasis on TSWV and Frankliniella occidentalis, the model tospovirus-thrips pathosystem. However, other virus-vector combinations are also of importance and where possible, comparisons are made between different viruses and thrips vectors. PMID:26340723

  2. The molecular biology of nairoviruses, an emerging group of tick-borne arboviruses.

    Science.gov (United States)

    Lasecka, Lidia; Baron, Michael D

    2014-06-01

    The nairoviruses are a rapidly emerging group of tick-borne bunyaviruses that includes pathogens of humans (Crimean-Congo hemorrhagic fever virus [CCHFV]) and livestock (Nairobi sheep disease virus [NSDV], also known as Ganjam virus), as well as a large number of viruses for which the normal vertebrate host has not been established. Studies on this group of viruses have been fairly limited, not least because CCHFV is a BSL4 human pathogen, restricting the number of labs able to study the live virus, while NSDV, although highly pathogenic in naive animals, is not seen as a threat in developed countries, making it a low priority. Nevertheless, recent years have seen significant progress in our understanding of the biology of these viruses, particularly that of CCHFV, and this article seeks to draw together our existing knowledge to generate an overall picture of their molecular biology, underlining areas of particular ignorance for future studies. PMID:24327094

  3. Crimean-Congo Hemorrhagic Fever Virus Subunit Vaccines Induce High Levels of Neutralizing Antibodies But No Protection in STAT1 Knockout Mice.

    Science.gov (United States)

    Kortekaas, Jeroen; Vloet, Rianka P M; McAuley, Alexander J; Shen, Xiaoli; Bosch, Berend Jan; de Vries, Laura; Moormann, Rob J M; Bente, Dennis A

    2015-12-01

    Crimean-Congo hemorrhagic fever virus is a tick-borne bunyavirus of the Nairovirus genus that causes hemorrhagic fever in humans with high case fatality. Here, we report the development of subunit vaccines and their efficacy in signal transducer and activator of transcription 1 (STAT1) knockout mice. Ectodomains of the structural glycoproteins Gn and Gc were produced using a Drosophila insect cell-based expression system. A single vaccination of STAT129 mice with adjuvanted Gn or Gc ectodomains induced neutralizing antibody responses, which were boosted by a second vaccination. Despite these antibody responses, mice were not protected from a CCHFV challenge infection. These results suggest that neutralizing antibodies against CCHFV do not correlate with protection of STAT1 knockout mice. PMID:26684523

  4. Single-particle cryo-electron microscopy of Rift Valley fever virus

    International Nuclear Information System (INIS)

    Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human and veterinary pathogen causing acute hepatitis in ruminants and has the potential to cause hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on the virus surface are highly symmetric and arranged on a T = 12 icosahedral lattice. Our RVFV MP-12 structure allowed clear identification of inter-capsomer contacts and definition of possible glycoprotein arrangements within capsomers. This structure provides a detailed model for phleboviruses, opens new avenues for high-resolution structural studies of the bunyavirus family, and aids the design of antiviral diagnostics and effective subunit vaccines.

  5. Immunological features underlying viral hemorrhagic fevers.

    Science.gov (United States)

    Messaoudi, Ilhem; Basler, Christopher F

    2015-10-01

    Several enveloped RNA viruses of the arenavirus, bunyavirus, filovirus and flavivirus families are associated with a syndrome known as viral hemorrhagic fever (VHF). VHF is characterized by fever, vascular leakage, coagulation defects and multi organ system failure. VHF is currently viewed as a disease precipitated by viral suppression of innate immunity, which promotes systemic virus replication and excessive proinflammatory cytokine responses that trigger the manifestations of severe disease. However, the mechanisms by which immune dysregulation contributes to disease remain poorly understood. Infection of nonhuman primates closely recapitulates human VHF, notably Ebola and yellow fever, thereby providing excellent models to better define the immunological basis for this syndrome. Here we review the current state of our knowledge and suggest future directions that will better define the immunological mechanisms underlying VHF. PMID:26163194

  6. Single-Molecule FISH Reveals Non-selective Packaging of Rift Valley Fever Virus Genome Segments.

    Science.gov (United States)

    Wichgers Schreur, Paul J; Kortekaas, Jeroen

    2016-08-01

    The bunyavirus genome comprises a small (S), medium (M), and large (L) RNA segment of negative polarity. Although genome segmentation confers evolutionary advantages by enabling genome reassortment events with related viruses, genome segmentation also complicates genome replication and packaging. Accumulating evidence suggests that genomes of viruses with eight or more genome segments are incorporated into virions by highly selective processes. Remarkably, little is known about the genome packaging process of the tri-segmented bunyaviruses. Here, we evaluated, by single-molecule RNA fluorescence in situ hybridization (FISH), the intracellular spatio-temporal distribution and replication kinetics of the Rift Valley fever virus (RVFV) genome and determined the segment composition of mature virions. The results reveal that the RVFV genome segments start to replicate near the site of infection before spreading and replicating throughout the cytoplasm followed by translocation to the virion assembly site at the Golgi network. Despite the average intracellular S, M and L genome segments approached a 1:1:1 ratio, major differences in genome segment ratios were observed among cells. We also observed a significant amount of cells lacking evidence of M-segment replication. Analysis of two-segmented replicons and four-segmented viruses subsequently confirmed the previous notion that Golgi recruitment is mediated by the Gn glycoprotein. The absence of colocalization of the different segments in the cytoplasm and the successful rescue of a tri-segmented variant with a codon shuffled M-segment suggested that inter-segment interactions are unlikely to drive the copackaging of the different segments into a single virion. The latter was confirmed by direct visualization of RNPs inside mature virions which showed that the majority of virions lack one or more genome segments. Altogether, this study suggests that RVFV genome packaging is a non-selective process. PMID:27548280

  7. Status of tick distribution and tick-borne pathogens in Jinan city%济南市蜱分布及带病毒状况调查

    Institute of Scientific and Technical Information of China (English)

    辛正; 王东; 杨国樑; 王永明; 彭文广; 李羡亭; 齐梅; 王蕾; 李殿香

    2015-01-01

    Objective To investigate the species, host, distribution and status of tick⁃borne pathogens in Jinan city. Methods The parasitic ticks were collected from the host skin by hand or tweezers and the free ticks were collected manually with white cloth from the grassland or shrubbery. Collected ticks were classified and tested for tick⁃borne pathogens. Results There were 614 and 108 ticks collected on 6 hosts and in 2 types of environment, respectively. Collected ticks were Haemaphysalis longicornis. There were 596 ticks collected on goats with proportion of 97.1%. About 53.3%goats carried with ticks and the average number of ticks per goat was about 6.7. The results were positive in RNA detection of new bunyavirus in 3 groups of tick and positive of rickettsia in one group. Positive ticks were collected from goats. Conclusion The dominant tick species was H. longicornis in Shandong province. The dominant host animal was goats raised outside. Some ticks may carry bunyavirus and rickettsia.%目的:调查济南市丘陵地区蜱的种类、宿主、分布及其带病毒情况。方法采用宿主体表捡拾法采集寄生蜱、人工布旗法采集游离蜱,并进行分类鉴定和病原体检测。结果在宿主动物和环境中分别捕获蜱614和108只,经鉴定均为长角血蜱。其中,羊体表捕获蜱596只,占调查宿主动物的97.1%。所有宿主动物中,羊携带蜱比例最高(53.3%),带蜱指数最高(6.7只/只或头)。在3组蜱样本中检测到新型布尼亚病毒核酸阳性,1组蜱样本中检测到立克次体阳性,4份样本均来自羊群。结论济南市的优势蜱种为长角血蜱;放养羊群是当地蜱主要宿主动物;部分蜱可能携带新型布尼亚病毒和立克次体。

  8. Jatobal virus antigenic characterization by ELISA and neutralization test using EIA as indicator, on tissue culture

    Directory of Open Access Journals (Sweden)

    Luiz Tadeu M. Figueiredo

    1988-06-01

    Full Text Available A virus antigenic characterization methodology using an indirect method of antibody detection ELISA with virus-infected cultured cells as antigen and a micro virus neutralisation test using EIA (NT-EIA as an aid to reading were used for antigenic characterization of Jatobal (BeAn 423380. Jatobal virus was characterized as a Bunyaviridae, Bunyavirus genus, Simbu serogroup virus. ELISA using infected cultured cells as antigen is a sensitive and reliable method for identification of viruses and has many advantages over conventional antibody capture ELISA's and other tests: it eliminates solid phase coating with virus and laborious antigen preparation; it permits screening of large numbers of virus antisera faster and more easily than by CF, HAI, or plaque reduction NT. ELISA and NT using EIA as an aid to reading can be applicable to viruses which do not produce cytopathogenic effect. Both techniques are applicable to identification of viruses which grow in mosquito cells.A caracterização antigênica do vírus Jatobal (BeAn 423380 foi efetuada utilizando uma técnica de ELISA para deteccão de anticorpos que utiliza culturas celulares infectadas como antígeno e um micro teste de neutralização para vírus que utiliza o método imunoenzimático como auxiliar para a leitura dos resultados (NT-EIA. O vírus Jatobal foi caracterizado como um Bunyaviridae, gênero Bunyavirus, pertencente ao sorogrupo Simbu. A técnica de ELISA, utilizando culturas celulares infectadas como antígeno, trata-se de método sensível e confiável na identificação de agentes virais, possuindo muitas vantagens sobre ELISA convencionais e outros testes: elimina a preparação laboriosa de antígenos para o revestimento em fase sólida; permite que se teste de forma mais rápida e fácil que por CF, HAI e neutralização por redução de plaques um grande número de antisoros de vírus. ELISA e NT-EIA podem ser utilizados para a classificação de vírus que não produzem

  9. Role of envelope glycoproteins in intracellular virus maturation

    International Nuclear Information System (INIS)

    The possible role viral glycoproteins in intracellular maturation was studied by using two different viruses, avian infectious bronchitis virus (IBV), a coronavirus, and Punta Toro virus (PTV), a bunyavirus. Using the antibiotic tunicamycin, which inhibits glycosylation of N-linked glycoproteins, it was shown that coronavirus particles are formed in the absence of glycosylation. Analysis of the protein composition of these particles indicated that they contain an unglycosylated form of the membrane-associated E1 glycoprotein but lack the E2 spike glycoprotein. A cDNA clone derived from the PTV M RNA genome segment, which encodes the G1 and G2 glycoproteins, was cloned into vaccinia virus. Studies by indirect immunofluorescence microscopy revealed that the glycoproteins synthesized from this recombinant were found to accumulate intracellularly at the Golgi complex, where virus budding usually takes place. Surface immunoprecipitation and 125I-protein A binding assays also demonstrated that a majority of the glycoproteins are retained intracellularly and are not transported to the cellular surface. The sequences which encode the G1 and G2 glycoproteins were independently cloned into vaccinia virus as well

  10. A Global Genomic Characterization of Nairoviruses Identifies Nine Discrete Genogroups with Distinctive Structural Characteristics and Host-Vector Associations.

    Science.gov (United States)

    Walker, Peter J; Widen, Steven G; Wood, Thomas G; Guzman, Hilda; Tesh, Robert B; Vasilakis, Nikolaos

    2016-05-01

    Nairoviruses are primarily tick-borne bunyaviruses, some of which are known to cause mild-to-severe febrile illness in humans or livestock. We describe the genome sequences of 11 poorly characterized nairoviruses that have ecological associations with either birds (Farallon, Punta Salinas, Sapphire II, Zirqa, Avalon, Clo Mor, Taggert, and Abu Hammad viruses), rodents (Qalyub and Bandia viruses), or camels (Dera Ghazi Khan virus). Global phylogenetic analyses of proteins encoded in the L, M, and S RNA segments of these and 20 other available nairovirus genomes identified nine well-supported genogroups (Nairobi sheep disease, Thiafora, Sakhalin, Keterah, Qalyub, Kasokero, Dera Ghazi Khan, Hughes, and Tamdy). Genogroup-specific structural variations were evident, particularly in the M segment encoding a polyprotein from which virion envelope glycoproteins (Gn and Gc) are generated by proteolytic processing. Structural variations include the extension, abbreviation, or absence sequences encoding an O-glycosylated mucin-like protein in the N-terminal domain, distinctive patterns of conserved cysteine residues in the GP38-like domain, insertion of sequences encoding a double-membrane-spanning protein (NSm) between the Gn and Gc domains, and the presence of an alternative long open reading frame encoding a viroporin-like transmembrane protein (Gx). We also observed strong genogroup-specific associations with categories of hosts and tick vectors. PMID:26903607

  11. PREVALENCE OF ARBOVIRUS ANTIBODIES AGAINST THE FAMILY Bunyaviridae IN WATER BUFFALOES

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    Alexandre Rosário Casseb

    2015-07-01

    Full Text Available The State of Pará comprises 26% of Brazilian Amazon region where a large diversity of arboviruses has been described. This study sought to assess the prevalence and distribution of haemagglutination-inhibition antibodies against antigens of nine different types of arbovirus of the Bunyaviridae family, where eight were Orthobunyavirus: Guaroa virus, Maguari virus, Tacaiuma virus, Utinga virus, Belem virus, Caraparu virus, Oropouche virus and Catu virus, and one Phlebovirus: Icoaraci virus in sera samples of water buffaloes in Pará State, Brazil. For all Arboviruses investigated there were antibodies, with the exception of Belem virus. Antibodies to Maguari virus were more prevalent (7.33%. The water buffaloes of the present study showed variable levels of antibodies in monotypic and heterotypic reactions that may indicate there are movements from most bunyavirus studied in domestic buffaloes in the state of Pará, and the Maguari virus presents the largest circulation. Therefore, further studies are needed to investigate the role of water buffalo in the maintenance and dispersal of arboviruses, as well as whether these viruses can cause disease in that species, especially in cases of birth defects and abortions.

  12. Evaluation of the eastern cottontail Sylvilagus floridanus as an amplifying vertebrate host for Cache Valley virus (Bunyaviridae) in Indiana.

    Science.gov (United States)

    Blackmore, Carina G M; Grimstad, Paul R

    2008-01-01

    To evaluate the importance of eastern cottontails (Sylvilagus floridanus) as amplifying hosts for Cache Valley virus (CVV), we tested hunter-provided blood samples from northern Indiana for specific neutralizing (N) antibodies against this mosquito-borne bunya-virus. Samples were collected during the winter of 1994-95. Two seronegative eastern cottontails, captured in July 1995, were also infected with CVV by subcutaneous inoculation, and two others were infected by allowing CVV-infected mosquitoes to feed on them. The results indicate that eastern cottontails probably are not important amplifying hosts for CVV. The prevalence of N antibodies against CVV was low (6.0%, n=82) among the hunter-killed animals. Low viremia (<1.8 log10 plaque-forming units/ml) of short duration (1-3 days) were seen in three of four experimentally infected eastern cottontails. The viremias were insufficient for infecting Coquillettidia perturbans, a mosquito species commonly found naturally infected with CVV. PMID:18263839

  13. Antiviral Chemistry & Chemotherapy's current antiviral agents FactFile 2008 (2nd edition): RNA viruses.

    Science.gov (United States)

    De Clercq, Erik; Field, Hugh J

    2008-01-01

    Among the RNA viruses, other than the retroviruses (that is, HIV), which are dealt with separately in the current FactFile, the most important targets for the development of antiviral agents at the moment are the orthomyxoviruses (that is, influenza), the hepaciviruses (that is, hepatitis C virus [HCV]) and, to a lesser extent, the picornaviruses. Although the uncoating inhibitors amantadine and rimantadine were the first known inhibitors of influenza A, the neuraminidase inhibitors oseltamivir, zanamivir and peramivir have now become the prime antiviral drugs for the treatment of influenza A and B virus infections. For HCV infections, standard treatment consists of the combination of pegylated interferon-alpha with ribavirin, but several other antivirals targeted at specific viral functions such as the HCV protease and/ or polymerase may be expected to soon take an important share of this important market. Still untapped is the potential of a variety of uncoating inhibitors, as well as protease and/or polymerase inhibitors against the wide spectrum of picornaviruses. While ribavirin has been available for 35 years as a broad-spectrum anti-RNA virus agent, relatively new and unexplored is favipiravir (T-705) accredited with activity against influenza as well as flaviviruses, bunyaviruses and arenaviruses. PMID:18727441

  14. The transient nature of Bunyamwera orthobunyavirus NSs protein expression: effects of increased stability of NSs protein on virus replication.

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    Ingeborg van Knippenberg

    Full Text Available The NSs proteins of bunyaviruses are the viral interferon antagonists, counteracting the host's antiviral response to infection. During high-multiplicity infection of cultured mammalian cells with Bunyamwera orthobunyavirus (BUNV, NSs is rapidly degraded after reaching peak levels of expression at 12hpi. Through the use of inhibitors this was shown to be the result of proteasomal degradation. A recombinant virus (rBUN4KR, in which all four lysine residues in NSs were replaced by arginine residues, expresses an NSs protein (NSs4KR that is resistant to degradation, confirming that degradation is lysine-dependent. However, despite repeated attempts, no direct ubiquitylation of NSs in infected cells could be demonstrated. This suggests that degradation of NSs, although lysine-dependent, may be achieved through an indirect mechanism. Infection of cultured mammalian cells or mice indicated no disadvantage for the virus in having a non-degradable NSs protein: in fact rBUN4KR had a slight growth advantage over wtBUNV in interferon-competent cells, presumably due to the increased and prolonged presence of NSs. In cultured mosquito cells there was no difference in growth between wild-type BUNV and rBUN4KR, but surprisingly NSs4KR was not stabilised compared to the wild-type NSs protein.

  15. The transient nature of Bunyamwera orthobunyavirus NSs protein expression: effects of increased stability of NSs protein on virus replication.

    Science.gov (United States)

    van Knippenberg, Ingeborg; Fragkoudis, Rennos; Elliott, Richard M

    2013-01-01

    The NSs proteins of bunyaviruses are the viral interferon antagonists, counteracting the host's antiviral response to infection. During high-multiplicity infection of cultured mammalian cells with Bunyamwera orthobunyavirus (BUNV), NSs is rapidly degraded after reaching peak levels of expression at 12hpi. Through the use of inhibitors this was shown to be the result of proteasomal degradation. A recombinant virus (rBUN4KR), in which all four lysine residues in NSs were replaced by arginine residues, expresses an NSs protein (NSs4KR) that is resistant to degradation, confirming that degradation is lysine-dependent. However, despite repeated attempts, no direct ubiquitylation of NSs in infected cells could be demonstrated. This suggests that degradation of NSs, although lysine-dependent, may be achieved through an indirect mechanism. Infection of cultured mammalian cells or mice indicated no disadvantage for the virus in having a non-degradable NSs protein: in fact rBUN4KR had a slight growth advantage over wtBUNV in interferon-competent cells, presumably due to the increased and prolonged presence of NSs. In cultured mosquito cells there was no difference in growth between wild-type BUNV and rBUN4KR, but surprisingly NSs4KR was not stabilised compared to the wild-type NSs protein. PMID:23667701

  16. Mapping of the interaction domains of the Crimean-Congo hemorrhagic fever virus nucleocapsid protein.

    Science.gov (United States)

    Levingston Macleod, Jesica M; Marmor, Hannah; García-Sastre, Adolfo; Frias-Staheli, Natalia

    2015-03-01

    Crimean-Congo hemorrhagic fever virus (CCHFV) is a member of the genus Nairovirus of the family Bunyaviridae, that can cause severe haemorrhagic fever in humans, with mortality rates above 30  %. CCHFV is the most widespread of the tick-borne human viruses and it is endemic in areas of central Asia, the Middle East, Africa and southern Europe. Its viral genome consists of three negative-sense RNA segments. The large segment (L) encodes a viral RNA-dependent RNA polymerase (L protein), the small segment (S) encodes the nucleocapsid protein (N protein) and the medium segment (M) encodes the envelope proteins. The N protein of bunyaviruses binds genomic RNA, forming the viral ribonucleoprotein (RNP) complex. The L protein interacts with these RNP structures, allowing the initiation of viral replication. The N protein also interacts with actin, although the regions and specific residues involved in these interactions have not yet been described. Here, by means of immunoprecipitation and immunofluorescence assays, we identified the regions within the CCHFV N protein implicated in homo-oligomerization and actin binding. We describe the interaction of the N protein with the CCHFV L protein, and identify the N- and C-terminal regions within the L protein that might be necessary for the formation of these N-L protein complexes. These results may guide the development of potent inhibitors of these complexes that could potentially block CCHFV replication. PMID:25389186

  17. Next-generation sequencing of southern African Crimean-Congo haemorrhagic fever virus isolates reveals a high frequency of M segment reassortment.

    Science.gov (United States)

    Goedhals, D; Bester, P A; Paweska, J T; Swanepoel, R; Burt, F J

    2014-09-01

    Crimean Congo haemorrhagic fever virus (CCHFV) is a bunyavirus with a single-stranded RNA genome consisting of three segments (S, M, L), coding for the nucleocapsid protein, envelope glycoproteins and RNA polymerase, respectively. To date only five complete genome sequences are available from southern African isolates. Complete genome sequences were generated for 10 southern African CCHFV isolates using next-generation sequencing techniques. The maximum-likelihood method was used to generate tree topologies for 15 southern African plus 26 geographically distinct complete sequences from GenBank. M segment reassortment was identified in 10/15 southern African isolates by incongruencies in grouping compared to the S and L segments. These reassortant M segments cluster with isolates from Asia/Middle East, while the S and L segments cluster with strains from South/West Africa. The CCHFV M segment shows a high level of genetic diversity, while the S and L segments appear to co-evolve. The reason for the high frequency of M segment reassortment is not known. It has previously been suggested that M segment reassortment results in a virus with high fitness but a clear role in increased pathogenicity has yet to be shown. PMID:24786748

  18. Mapping of the interaction domains of the Crimean–Congo hemorrhagic fever virus nucleocapsid protein

    Science.gov (United States)

    Macleod, Jesica M. Levingston; Marmor, Hannah; Frias-Staheli, Natalia

    2015-01-01

    Crimean–Congo hemorrhagic fever virus (CCHFV) is a member of the genus Nairovirus of the family Bunyaviridae, that can cause severe haemorrhagic fever in humans, with mortality rates above 30 %. CCHFV is the most widespread of the tick-borne human viruses and it is endemic in areas of central Asia, the Middle East, Africa and southern Europe. Its viral genome consists of three negative-sense RNA segments. The large segment (L) encodes a viral RNA-dependent RNA polymerase (L protein), the small segment (S) encodes the nucleocapsid protein (N protein) and the medium segment (M) encodes the envelope proteins. The N protein of bunyaviruses binds genomic RNA, forming the viral ribonucleoprotein (RNP) complex. The L protein interacts with these RNP structures, allowing the initiation of viral replication. The N protein also interacts with actin, although the regions and specific residues involved in these interactions have not yet been described. Here, by means of immunoprecipitation and immunofluorescence assays, we identified the regions within the CCHFV N protein implicated in homo-oligomerization and actin binding. We describe the interaction of the N protein with the CCHFV L protein, and identify the N- and C-terminal regions within the L protein that might be necessary for the formation of these N–L protein complexes. These results may guide the development of potent inhibitors of these complexes that could potentially block CCHFV replication. PMID:25389186

  19. Genomic Characterization of Yogue, Kasokero, Issyk-Kul, Keterah, Gossas, and Thiafora Viruses: Nairoviruses Naturally Infecting Bats, Shrews, and Ticks.

    Science.gov (United States)

    Walker, Peter J; Widen, Steven G; Firth, Cadhla; Blasdell, Kim R; Wood, Thomas G; Travassos da Rosa, Amelia P A; Guzman, Hilda; Tesh, Robert B; Vasilakis, Nikos

    2015-11-01

    The genus Nairovirus of arthropod-borne bunyaviruses includes the important emerging human pathogen, Crimean-Congo hemorrhagic fever virus (CCHFV), as well as Nairobi sheep disease virus and many other poorly described viruses isolated from mammals, birds, and ticks. Here, we report genome sequence analysis of six nairoviruses: Thiafora virus (TFAV) that was isolated from a shrew in Senegal; Yogue (YOGV), Kasokero (KKOV), and Gossas (GOSV) viruses isolated from bats in Senegal and Uganda; Issyk-Kul virus (IKV) isolated from bats in Kyrgyzstan; and Keterah virus (KTRV) isolated from ticks infesting a bat in Malaysia. The S, M, and L genome segments of each virus were found to encode proteins corresponding to the nucleoprotein, polyglycoprotein, and polymerase protein of CCHFV. However, as observed in Leopards Hill virus (LPHV) and Erve virus (ERVV), polyglycoproteins encoded in the M segment lack sequences encoding the double-membrane-spanning CCHFV NSm protein. Amino acid sequence identities, complement-fixation tests, and phylogenetic analysis indicated that these viruses cluster into three groups comprising KKOV, YOGV, and LPHV from bats of the suborder Yingochiroptera; KTRV, IKV, and GOSV from bats of the suborder Yangochiroptera; and TFAV and ERVV from shrews (Soricomorpha: Soricidae). This reflects clade-specific host and vector associations that extend across the genus. PMID:26324724

  20. Critical epitopes in the nucleocapsid protein of SFTS virus recognized by a panel of SFTS patients derived human monoclonal antibodies.

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    Li Yu

    Full Text Available BACKGROUND: SFTS virus (SFTSV is a newly discovered pathogen to cause severe fever with thrombocytopenia syndrome (SFTS in human. Successful control of SFTSV epidemic requires better understanding of the antigen target in humoral immune responses to the new bunyavirus infection. METHODOLOGY/PRINCIPAL FINDINGS: We have generated a combinatorial Fab antibody phage library from two SFTS patients recovered from SFTSV infection. To date, 94 unique human antibodies have been generated and characterized from over 1200 Fab antibody clones obtained by screening the library with SFTS purified virions. All those monoclonal antibodies (MAbs recognized the nucleocapsid (N protein of SFTSV while none of them were reactive to the viral glycoproteins Gn or Gc. Furthermore, over screening 1000 mouse monoclonal antibody clones derived from SFTSV virions immunization, 462 clones reacted with N protein, while only 16 clones were reactive to glycoprotein. Furthermore, epitope mapping of SFTSV N protein was performed through molecular simulation, site mutation and competitive ELISA, and we found that at least 4 distinct antigenic epitopes within N protein were recognized by those human and mouse MAbs, in particular mutation of Glu10 to Ala10 abolished or significantly reduced the binding activity of nearly most SFTS patients derived MAbs. CONCLUSIONS/SIGNIFICANCE: The large number of human recombinant MAbs derived from SFTS patients recognized the viral N protein indicated the important role of the N protein in humoral responses to SFTSV infection, and the critical epitopes we defined in this study provided molecular basis for detection and diagnosis of SFTSV infection.

  1. Bovine Lactoferrin Inhibits Toscana Virus Infection by Binding to Heparan Sulphate

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    Agostina Pietrantoni

    2015-01-01

    Full Text Available Toscana virus is an emerging sandfly-borne bunyavirus in Mediterranean Europe responsible for neurological diseases in humans. It accounts for about 80% of paediatric meningitis cases during the summer. Despite the important impact of Toscana virus infection-associated disease on human health, currently approved vaccines or effective antiviral treatments are not available. In this research, we have analyzed the effect of bovine lactoferrin, a bi-globular iron-binding glycoprotein with potent antimicrobial and immunomodulatory activities, on Toscana virus infection in vitro. Our results showed that lactoferrin was capable of inhibiting Toscana virus replication in a dose-dependent manner. Results obtained when lactoferrin was added to the cells during different phases of viral infection showed that lactoferrin was able to prevent viral replication when added during the viral adsorption step or during the entire cycle of virus infection, demonstrating that its action takes place in an early phase of viral infection. In particular, our results demonstrated that the anti-Toscana virus action of lactoferrin took place on virus attachment to the cell membrane, mainly through a competition for common glycosaminoglycan receptors. These findings provide further insights on the antiviral activity of bovine lactoferrin.

  2. Bovine lactoferrin inhibits Toscana virus infection by binding to heparan sulphate.

    Science.gov (United States)

    Pietrantoni, Agostina; Fortuna, Claudia; Remoli, Maria Elena; Ciufolini, Maria Grazia; Superti, Fabiana

    2015-02-01

    Toscana virus is an emerging sandfly-borne bunyavirus in Mediterranean Europe responsible for neurological diseases in humans. It accounts for about 80% of paediatric meningitis cases during the summer. Despite the important impact of Toscana virus infection-associated disease on human health, currently approved vaccines or effective antiviral treatments are not available. In this research, we have analyzed the effect of bovine lactoferrin, a bi-globular iron-binding glycoprotein with potent antimicrobial and immunomodulatory activities, on Toscana virus infection in vitro. Our results showed that lactoferrin was capable of inhibiting Toscana virus replication in a dose-dependent manner. Results obtained when lactoferrin was added to the cells during different phases of viral infection showed that lactoferrin was able to prevent viral replication when added during the viral adsorption step or during the entire cycle of virus infection, demonstrating that its action takes place in an early phase of viral infection. In particular, our results demonstrated that the anti-Toscana virus action of lactoferrin took place on virus attachment to the cell membrane, mainly through a competition for common glycosaminoglycan receptors. These findings provide further insights on the antiviral activity of bovine lactoferrin. PMID:25643293

  3. A preliminary study of viral metagenomics of French bat species in contact with humans: identification of new mammalian viruses.

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    Laurent Dacheux

    Full Text Available The prediction of viral zoonosis epidemics has become a major public health issue. A profound understanding of the viral population in key animal species acting as reservoirs represents an important step towards this goal. Bats harbor diverse viruses, some of which are of particular interest because they cause severe human diseases. However, little is known about the diversity of the global population of viruses found in bats (virome. We determined the viral diversity of five different French insectivorous bat species (nine specimens in total in close contact with humans. Sequence-independent amplification, high-throughput sequencing with Illumina technology and a dedicated bioinformatics analysis pipeline were used on pooled tissues (brain, liver and lungs. Comparisons of the sequences of contigs and unassembled reads provided a global taxonomic distribution of virus-related sequences for each sample, highlighting differences both within and between bat species. Many viral families were present in these viromes, including viruses known to infect bacteria, plants/fungi, insects or vertebrates, the most relevant being those infecting mammals (Retroviridae, Herpesviridae, Bunyaviridae, Poxviridae, Flaviviridae, Reoviridae, Bornaviridae, Picobirnaviridae. In particular, we detected several new mammalian viruses, including rotaviruses, gammaretroviruses, bornaviruses and bunyaviruses with the identification of the first bat nairovirus. These observations demonstrate that bats naturally harbor viruses from many different families, most of which infect mammals. They may therefore constitute a major reservoir of viral diversity that should be analyzed carefully, to determine the role played by bats in the spread of zoonotic viral infections.

  4. Microarrays - new possibilities for detecting biological factors hazardous for humans and animals, and for use in environmental protection.

    Science.gov (United States)

    Mirski, Tomasz; Bartoszcze, Michał; Bielawska-Drózd, Agata; Gryko, Romuald; Niemcewicz, Marcin; Kocik, Janusz; Chomiczewski, Krzysztof

    2016-03-01

    Both the known biological agents that cause infectious diseases, as well as modified (ABF-Advanced Biological Factors) or new, emerging agents pose a significant diagnostic problem using previously applied methods, both classical, as well as based on molecular biology methods. The latter, such as PCR and real-time PCR, have significant limitations, both quantitative (low capacity), and qualitative (limited number of targets). The article discusses the results of studies on using the microarray method for the identification of viruses (e.g. Orthopoxvirus group, noroviruses, influenza A and B viruses, rhino- and enteroviruses responsible for the FRI (Febrile Respiratory Illness), European bunyaviruses, and SARS-causing viruses), and bacteria (Mycobacterium spp., Yersinia spp., Campylobacter spp., Streptococcus pneumoniae, Salmonella typhi, Salmonella enterica, Staphylococcus aureus, Neisseria meningitidis, Clostridium difficile , Helicobacter pylori), including multiple antibiotic-resistant strains. The method allows for the serotyping and genotyping of bacteria, and is useful in the diagnosis of genetically modified agents. It allows the testing of thousands of genes in one experiment. In addition to diagnosis, it is applicable for gene expression studies, analysis of the function of genes, microorganisms virulence, and allows the detection of even single mutations. The possibility of its operational application in epidemiological surveillance, and in the detection of disease outbreak agents is demonstrated. PMID:27007515

  5. Differential Use of the C-Type Lectins L-SIGN and DC-SIGN for Phlebovirus Endocytosis.

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    Léger, Psylvia; Tetard, Marilou; Youness, Berthe; Cordes, Nicole; Rouxel, Ronan N; Flamand, Marie; Lozach, Pierre-Yves

    2016-06-01

    Bunyaviruses represent a growing threat to humans and livestock globally. The receptors, cellular factors and endocytic pathways used by these emerging pathogens to infect cells remain largely unidentified and poorly characterized. DC-SIGN is a C-type lectin highly expressed on dermal dendritic cells that has been found to act as an authentic entry receptor for many phleboviruses (Bunyaviridae), including Rift Valley fever virus (RVFV), Toscana virus (TOSV) and Uukuniemi virus (UUKV). We found that these phleboviruses can exploit another C-type lectin, L-SIGN, for infection. L-SIGN shares 77% sequence homology with DC-SIGN and is expressed on liver sinusoidal endothelial cells. L-SIGN is required for UUKV binding but not for virus internalization. An endocytosis-defective mutant of L-SIGN was still able to mediate virus uptake and infection, indicating that L-SIGN acts as an attachment receptor for phleboviruses rather than an endocytic receptor. Our results point out a fundamental difference in the use of the C-type lectins L-SIGN and DC-SIGN by UUKV to enter cells, although both proteins are closely related in terms of molecular structure and biological function. This study sheds new light on the molecular mechanisms by which phleboviruses target the liver and also highlights the added complexity in virus-receptor interactions beyond attachment. PMID:26990254

  6. Conserved Endonuclease Function of Hantavirus L Polymerase

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    Sylvia Rothenberger

    2016-05-01

    Full Text Available Hantaviruses are important emerging pathogens belonging to the Bunyaviridae family. Like other segmented negative strand RNA viruses, the RNA-dependent RNA polymerase (RdRp also known as L protein of hantaviruses lacks an intrinsic “capping activity”. Hantaviruses therefore employ a “cap snatching” strategy acquiring short 5′ RNA sequences bearing 5′cap structures by endonucleolytic cleavage from host cell transcripts. The viral endonuclease activity implicated in cap snatching of hantaviruses has been mapped to the N-terminal domain of the L protein. Using a combination of molecular modeling and structure–function analysis we confirm and extend these findings providing evidence for high conservation of the L endonuclease between Old and New World hantaviruses. Recombinant hantavirus L endonuclease showed catalytic activity and a defined cation preference shared by other viral endonucleases. Based on the previously reported remarkably high activity of hantavirus L endonuclease, we established a cell-based assay for the hantavirus endonuclase function. The robustness of the assay and its high-throughput compatible format makes it suitable for small molecule drug screens to identify novel inhibitors of hantavirus endonuclease. Based on the high degree of similarity to RdRp endonucleases, some candidate inhibitors may be broadly active against hantaviruses and other emerging human pathogenic Bunyaviruses.

  7. Conserved Endonuclease Function of Hantavirus L Polymerase.

    Science.gov (United States)

    Rothenberger, Sylvia; Torriani, Giulia; Johansson, Maria U; Kunz, Stefan; Engler, Olivier

    2016-01-01

    Hantaviruses are important emerging pathogens belonging to the Bunyaviridae family. Like other segmented negative strand RNA viruses, the RNA-dependent RNA polymerase (RdRp) also known as L protein of hantaviruses lacks an intrinsic "capping activity". Hantaviruses therefore employ a "cap snatching" strategy acquiring short 5' RNA sequences bearing 5'cap structures by endonucleolytic cleavage from host cell transcripts. The viral endonuclease activity implicated in cap snatching of hantaviruses has been mapped to the N-terminal domain of the L protein. Using a combination of molecular modeling and structure-function analysis we confirm and extend these findings providing evidence for high conservation of the L endonuclease between Old and New World hantaviruses. Recombinant hantavirus L endonuclease showed catalytic activity and a defined cation preference shared by other viral endonucleases. Based on the previously reported remarkably high activity of hantavirus L endonuclease, we established a cell-based assay for the hantavirus endonuclase function. The robustness of the assay and its high-throughput compatible format makes it suitable for small molecule drug screens to identify novel inhibitors of hantavirus endonuclease. Based on the high degree of similarity to RdRp endonucleases, some candidate inhibitors may be broadly active against hantaviruses and other emerging human pathogenic Bunyaviruses. PMID:27144576

  8. Genetic and Phylogenetic Characterization of Tataguine and Witwatersrand Viruses and Other Orthobunyaviruses of the Anopheles A, Capim, Guamá, Koongol, Mapputta, Tete, and Turlock Serogroups.

    Science.gov (United States)

    Shchetinin, Alexey M; Lvov, Dmitry K; Deriabin, Petr G; Botikov, Andrey G; Gitelman, Asya K; Kuhn, Jens H; Alkhovsky, Sergey V

    2015-11-01

    The family Bunyaviridae has more than 530 members that are distributed among five genera or remain to be classified. The genus Orthobunyavirus is the most diverse bunyaviral genus with more than 220 viruses that have been assigned to more than 18 serogroups based on serological cross-reactions and limited molecular-biological characterization. Sequence information for all three orthobunyaviral genome segments is only available for viruses belonging to the Bunyamwera, Bwamba/Pongola, California encephalitis, Gamboa, Group C, Mapputta, Nyando, and Simbu serogroups. Here we present coding-complete sequences for all three genome segments of 15 orthobunyaviruses belonging to the Anopheles A, Capim, Guamá, Kongool, Tete, and Turlock serogroups, and of two unclassified bunyaviruses previously not known to be orthobunyaviruses (Tataguine and Witwatersrand viruses). Using those sequence data, we established the most comprehensive phylogeny of the Orthobunyavirus genus to date, now covering 15 serogroups. Our results emphasize the high genetic diversity of orthobunyaviruses and reveal that the presence of the small nonstructural protein (NSs)-encoding open reading frame is not as common in orthobunyavirus genomes as previously thought. PMID:26610546

  9. Epidemia de febre do Oropouche em Serra Pelada, município de Curionópolis, Pará, 1994

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    Amélia P.A.T. Rosa

    1996-12-01

    Full Text Available No final de novembro de 1994, o Instituto Evandro Chagas (IEC, Belém, Pará, foi notificado de um surto de doença febril na população do garimpo de Serra Pelada, município de Curionôpolis (5°35'S; 49°30'W, no Estado do Pará. Vinte amostras de soro de pessoas, com hemoscopia negativa para tnalária, foram recebidas para esclarecimento diagnóstico. Estudos laboratoriais comprovaram que os casos eram devido ao vírus Oropouche (grupo Simbu. gênero Bunyavirus, família Bunyaviridae. Esses achados, induziram d ida de um grupo de técnicos para realização de investigações ecoepidemíológicas entre 8 e 22 de dezembro. Foram coletadas 296 amostras de sangue, de 73 grupos familiares, sendo 54 para pequisa de vírus (casos febris e 242para sorologia, bem como, procedeu-se a coleta de artrópodes hematófagos. As amostras para pesquisa de vírus foram inoculadas em camundongos recém-nascidos e os soros testados por inibição da hemaglutinação (1H e MAC ELISA. Foram isoladas dez amostras do vírus Oropouche e obtidas seis soroconversões. Ademais, 245 (82,8% amostras foram positivas por sorologia e 71 (97,3% grupos familiares apresentaram pelo menos um membro positivo. Considerando a elevada positividade de anticoipos IH e IgM específica para Oropouche na população de Serra Pelada, concluímos que a epidemia foi extensa e apresentou taxa de ataque em torno de 83%, que correspondeu a infecção de cerca de 5.000 pessoas.In the final of November 1994, an outbreak of a febrile disease was observed in the Serra Pelada gold mine (5°35'S; 49°30'W in the Southeast region of Parã State. Twenty samples were collected and sent to the laboratory of Arbovirus of Instituto Evandro Chagas. The tests showed that the disease was caused by Oropouche virus (Bunyaviridae, Bunyavirus, Simbu serological group. Between 8-22 December 296 serum samples mere taken (54 from febrile patients, 16 paired samples and 242 from contacts and convalescent patients

  10. Detection of Severe Fever with Thrombocytopenia Syndrome Virus from Wild Animals and Ixodidae Ticks in the Republic of Korea.

    Science.gov (United States)

    Oh, Sung-Suck; Chae, Jeong-Byoung; Kang, Jun-Gu; Kim, Heung-Chul; Chong, Sung-Tae; Shin, Jeong-Hwa; Hur, Moon-Suk; Suh, Jae-Hwa; Oh, Myoung-Don; Jeong, Soo-Myoung; Shin, Nam-Shik; Choi, Kyoung-Seong; Chae, Joon-Seok

    2016-06-01

    Severe fever with thrombocytopenia syndrome (SFTS) is caused by SFTS virus (SFTSV), a novel bunyavirus reported to be endemic to central-northeastern China, southern Japan, and the Republic of Korea (ROK). To investigate SFTSV infections, we collected serum samples and ticks from wild animals. Using serum samples and ticks, SFTSV-specific genes were amplified by one-step RT-PCR and nested PCR and sequenced. Indirect immunofluorescence assay (IFA) was performed to analyze virus-specific antibody levels in wild animals. Serum samples were collected from a total of 91 animals: 21 Korean water deer (KWD), 3 Siberian roe deer, 5 gorals, 7 raccoon dogs, 54 wild boars (WBs), and 1 carrion crow. The SFTSV infection rate in wild animals was 3.30% (3 of 91 animals: 1 KWD and 2 WBs). The seropositive rate was 6.59% (6 of 91 animals: 5 KWD and 1 WB). A total of 891 ticks (3 species) were collected from 65 wild animals (9 species). Of the attached tick species, Haemaphysalis longicornis (74.86%) was the most abundant, followed by Haemaphysalis flava (20.20%) and Ixodes nipponensis (4.94%). The average minimum infection rate (MIR) of SFTSV in ticks was 4.98%. The MIRs of H. longicornis, H. flava, and I. nipponensis were 4.51%, 2.22%, and 22.73%, respectively. The MIRs of larvae, nymphs, and adult ticks were 0.68%, 6.88%, and 5.53%, respectively. In addition, the MIRs of fed and unfed ticks were 4.67% and 4.96%, respectively. We detected a low SFTSV infection rate in wild animals, no differences in SFTSV infection rate with respect to bloodsucking in ticks, and SFTSV infection for all developmental stages of ticks. This is the first report describing the detection of SFTSV in wild animals in the ROK. PMID:27043361

  11. Association of the Serum Angiotensin II Level with Disease Severity in Severe Fever with Thrombocytopenia Syndrome Patients.

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    Cheng, Jiamei; Li, Huiyu; Jie, Shenghua

    2016-01-01

    Objective Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by a novel Bunyavirus. Recent data suggest that the physiological balance of multiple proinflammatory cytokines is substantially changed in cases of severe fever with thrombocytopenia syndrome virus (SFTSV) infection, and the inflammatory response probably plays an important role in disease progression. Angiotensin II is an important active substance of the renin-angiotensin system, and studies have demonstrated that angiotensin II is involved in key events in the inflammatory process and can regulate inflammatory cell responses. Methods In order to elucidate the role of angiotensin II in the pathogenesis of SFTS, we collected serum samples from SFTS patients in the acute or convalescent phase and tested the angiotensin II levels using an enzyme-linked immunosorbent assay as well as SFTSV viral RNA with real-time reverse-transcriptase polymerase chain reaction. Furthermore, we explored possible correlations between the angiotensin II levels and clinical parameters in SFTS patients. Results Our data showed that the serum level of angiotensin II was significantly increased in the acute phase compared with that seen in the convalescent phase and the healthy controls, while there were no significant differences between the convalescent cases and healthy controls (p>0.05). A correlation analysis demonstrated that the level of angiotensin II positively correlated with the SFTS viral RNA load. The angiotensin II levels were also found to be correlated with clinical parameters indicating impairments in organ functions. Moreover, we also found that the angiotensin II levels were significantly increased in the severe cases versus the non-severe cases (p<0.001). Conclusion The serum angiotensin II levels in SFTS patients may be used to stratify the disease severity and are possibly predictive of disease outcomes. PMID:27086801

  12. Inhibition of Hazara nairovirus replication by small interfering RNAs and their combination with ribavirin

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    Crance Jean-Marc

    2011-05-01

    Full Text Available Abstract Background The genus Nairovirus in the family Bunyaviridae contains 34 tick-borne viruses classified into seven serogroups. Hazara virus (HAZV belongs to the Crimean-Congo hemorrhagic fever (CCHF serogroup that also includes CCHF virus (CCHFV a major pathogen for humans. HAZV is an interesting model to study CCHFV due to a close serological and phylogenetical relationship and a classification which allows handling in a BSL2 laboratory. Nairoviruses are characterized by a tripartite negative-sense single stranded RNA genome (named L, M and S segments that encode the RNA polymerase, the Gn-Gc glycoproteins and the nucleoprotein (NP, respectively. Currently, there are neither vaccines nor effective therapies for the treatment of any bunyavirus infection in humans. In this study we report, for the first time, the use of RNA interference (RNAi as an approach to inhibit nairovirus replication. Results Chemically synthesized siRNAs were designed to target the mRNA produced by the three genomic segments. We first demonstrated that the siRNAs targeting the NP mRNA displayed a stronger antiviral effect than those complementary to the L and M transcripts in A549 cells. We further characterized the two most efficient siRNAs showing, that the induced inhibition is specific and associated with a decrease in NP synthesis during HAZV infection. Furthermore, both siRNAs depicted an antiviral activity when used before and after HAZV infection. We next showed that HAZV was sensitive to ribavirin which is also known to inhibit CCHFV. Finally, we demonstrated the additive or synergistic antiviral effect of siRNAs used in combination with ribavirin. Conclusions Our study highlights the interest of using RNAi (alone or in combination with ribavirin to treat nairovirus infection. This approach has to be considered for the development of future antiviral compounds targeting CCHFV, the most pathogenic nairovirus.

  13. Crimean-Congo hemorrhagic fever virus entry into host cells occurs through the multivesicular body and requires ESCRT regulators.

    Science.gov (United States)

    Shtanko, Olena; Nikitina, Raisa A; Altuntas, Cengiz Z; Chepurnov, Alexander A; Davey, Robert A

    2014-09-01

    Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne bunyavirus causing outbreaks of severe disease in humans, with a fatality rate approaching 30%. There are no widely accepted therapeutics available to prevent or treat the disease. CCHFV enters host cells through clathrin-mediated endocytosis and is subsequently transported to an acidified compartment where the fusion of virus envelope with cellular membranes takes place. To better understand the uptake pathway, we sought to identify host factors controlling CCHFV transport through the cell. We demonstrate that after passing through early endosomes in a Rab5-dependent manner, CCHFV is delivered to multivesicular bodies (MVBs). Virus particles localized to MVBs approximately 1 hour after infection and affected the distribution of the organelle within cells. Interestingly, blocking Rab7 activity had no effect on association of the virus with MVBs. Productive virus infection depended on phosphatidylinositol 3-kinase (PI3K) activity, which meditates the formation of functional MVBs. Silencing Tsg101, Vps24, Vps4B, or Alix/Aip1, components of the endosomal sorting complex required for transport (ESCRT) pathway controlling MVB biogenesis, inhibited infection of wild-type virus as well as a novel pseudotyped vesicular stomatitis virus (VSV) bearing CCHFV glycoprotein, supporting a role for the MVB pathway in CCHFV entry. We further demonstrate that blocking transport out of MVBs still allowed virus entry while preventing vesicular acidification, required for membrane fusion, trapped virions in the MVBs. These findings suggest that MVBs are necessary for infection and are the sites of virus-endosome membrane fusion. PMID:25233119

  14. Simultaneous detection of IgG antibodies associated with viral hemorrhagic fever by a multiplexed Luminex-based immunoassay.

    Science.gov (United States)

    Wu, Wei; Zhang, Shuo; Qu, Jing; Zhang, Quanfu; Li, Chuan; Li, Jiandong; Jin, Cong; Liang, Mifang; Li, Dexin

    2014-07-17

    Viral hemorrhagic fevers (VHFs) are worldwide diseases caused by several kinds of viruses. With the emergence of new viruses, advanced diagnostic methods are urgently needed for identification of VHFs. Based on Luminex xMAP technology, a rapid, sensitive, multi-pathogen and high-throughput method which could simultaneously detect hemorrhagic fever viruses (HFVs) specific IgG antibodies was developed. Recombinant antigens of nine HFVs including Hantaan virus (HTNV), Seoul virus (SEOV), Puumala virus (PUUV), Andes virus (ANDV), Sin Nombre virus (SNV), Crimean-Congo hemorrhagic fever virus (CCHFV), Rift Valley fever virus (RVFV), Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) and dengue virus (DENV) were produced and purified from a prokaryotic expression system and the influence of the coupling amount was investigated. Cross-reactions among antigens and their rabbit immune sera were evaluated. Serum samples collected from 51 laboratory confirmed hemorrhagic fever with renal syndrome (HFRS) patients, 43 confirmed SFTS patients and 88 healthy donors were analyzed. Results showed that recombinant nucleocapsid protein of the five viruses belonging to the genus Hantavirus, had serological cross-reactivity with their corresponding rabbit immune sera, but not apparent with immune sera of other four viruses. Evaluation of this new method with clinical serum samples showed 98.04% diagnostic sensitivity for HFRS, 90.70% for SFTS detection and the specificity was ranging from 66.67% to 100.00%. The multiplexed Luminex-based immunoassay has firstly been established in our study, which provides a potentially reliable diagnostic tool for IgG antibody detection of VHFs. PMID:24631566

  15. Antibodies to the core proteins of Nairobi sheep disease virus/Ganjam virus reveal details of the distribution of the proteins in infected cells and tissues.

    Science.gov (United States)

    Lasecka, Lidia; Bin-Tarif, Abdelghani; Bridgen, Anne; Juleff, Nicholas; Waters, Ryan A; Baron, Michael D

    2015-01-01

    Nairobi sheep disease virus (NSDV; also called Ganjam virus in India) is a bunyavirus of the genus Nairovirus. It causes a haemorrhagic gastroenteritis in sheep and goats with mortality up to 90%. The virus is closely related to the human pathogen Crimean-Congo haemorrhagic fever virus (CCHFV). Little is currently known about the biology of NSDV. We have generated specific antibodies against the virus nucleocapsid protein (N) and polymerase (L) and used these to characterise NSDV in infected cells and to study its distribution during infection in a natural host. Due to its large size and the presence of a papain-like protease (the OTU-like domain) it has been suggested that the L protein of nairoviruses undergoes an autoproteolytic cleavage into polymerase and one or more accessory proteins. Specific antibodies which recognise either the N-terminus or the C-terminus of the NSDV L protein showed no evidence of L protein cleavage in NSDV-infected cells. Using the specific anti-N and anti-L antibodies, it was found that these viral proteins do not fully colocalise in infected cells; the N protein accumulated near the Golgi at early stages of infection while the L protein was distributed throughout the cytoplasm, further supporting the multifunctional nature of the L protein. These antibodies also allowed us to gain information about the organs and cell types targeted by the virus in vivo. We could detect NSDV in cryosections prepared from various tissues collected post-mortem from experimentally inoculated animals; the virus was found in the mucosal lining of the small and large intestine, in the lungs, and in mesenteric lymph nodes (MLN), where NSDV appeared to target monocytes and/or macrophages. PMID:25905707

  16. Establishment of cell line from embryonic tissue of Pipistrellus ceylonicus bat species from India & its susceptibility to different viruses

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    Devendra T Mourya

    2013-01-01

    Full Text Available Background & objectives: Pipistrellus ceylonicus bat species is widely distributed in South Asia, with additional populations recorded in China and Southeast Asia. Bats are the natural reservoir hosts for a number of emerging zoonotic diseases. Attempts to isolate bat-borne viruses in various terrestrial mammalian cell lines have sometimes been unsuccessful. The bat cell lines are useful in isolation and propagation of many of the viruses harboured by bats. New stable bat cell lines are needed to help such investigations and to assist in the study of bat immunology and virus-host interactions. In this study we made an attempt to develop a cell line from P. ceylonicus bats. Methods: An effort was made to establish cell line from embryo of P. ceylonicus species of bat after seeding to Dulbecco′s modified eagle medium (DMEM supplemented with 10 per cent foetal bovine serum; a primary cell line was established and designated as NIV-BtEPC. Mitochondrial DNA profile analysis was done using cyt-b and ND-1 gene sequences from the cell line. Phylogenetic tree was constructed using neighbour-joining algorithm for cyt-b and ND-1 genes with 1000-bootstrap replicates. Results: NIV-BtEPC cell line was susceptible to Chandipura (CHPV and novel adenovirus (BtAdv-RLM isolated from Rousettus leschenaulti from India but did not support multiplication of a number of Bunyaviruses, Alphaviruses and Flavivirus. This might be useful for isolation of a range of viruses and investigation of unknown aetiological agents. Interpretation & conclusions: In this study, a new bat cell line was developed from P. ceylonicus. This cell line was successfully tested for the susceptibility to Chandipura and BtAdv-RLM virus isolated from bats. The approach developed and optimised in this study may be applicable to the other species of bats and this established cell line can be used to facilitate virus isolation and basic research into virus-host interaction.

  17. Epidemiological and clinical characteristics of severe fever with thrombocytopenia syndrome (SFTS) in China: an integrated data analysis.

    Science.gov (United States)

    Guo, C-T; Lu, Q-B; Ding, S-J; Hu, C-Y; Hu, J-G; Wo, Y; Fan, Y-D; Wang, X-J; Qin, S-L; Cui, N; Yang, Z-D; Zhang, X-A; Liu, W; Cao, W-C

    2016-04-01

    Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease that was caused by a novel bunyavirus, SFTSV. The study aimed to disclose the epidemiological and clinical characteristics of SFTSV infection in China so far. An integrated clinical database comprising 1920 SFTS patients was constructed by combining first-hand clinical information collected from SFTS sentinel hospitals (n = 1159) and extracted data (n = 761) from published literature. The considered variables comprised clinical manifestations, routine laboratory tests of acute infection, hospitalization duration and disease outcome. SFTSV-IgG data from 19 119 healthy subjects were extracted from the published papers. The key clinical variables, case-fatality rate (CFR) and seroprevalence were estimated by meta-analysis. The most commonly seen clinical manifestations of SFTSV infection were fever, anorexia, myalgia, chill and lymphadenopathy. The major laboratory findings were elevated lactate dehydrogenase, aminotransferase, followed by thrombocytopenia, lymphocytopenia, elevated alanine transaminase and creatine kinase. A CFR of 12·2% was estimated, significantly higher than that obtained from national reporting data, but showing no geographical difference. In our paper, the mortality rate was about 1·9 parts per million. Older age and longer delay to hospitalization were significantly associated with fatal outcome. A pooled seroprevalence of 3·0% was obtained, which increased with age, while comparable for gender. This study represents a clinical characterization on the largest group of SFTS patients up to now. A higher than expected CFR was obtained. A wider spectrum of clinical index was suggested to be used to identify SFTSV infection, while the useful predictor for fatal outcome was found to be restricted. PMID:26542444

  18. High-resolution structure of the N-terminal endonuclease domain of the Lassa virus L polymerase in complex with magnesium ions.

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    Gregor D Wallat

    Full Text Available Lassa virus (LASV causes deadly hemorrhagic fever disease for which there are no vaccines and limited treatments. LASV-encoded L polymerase is required for viral RNA replication and transcription. The functional domains of L-a large protein of 2218 amino acid residues-are largely undefined, except for the centrally located RNA-dependent RNA polymerase (RdRP motif. Recent structural and functional analyses of the N-terminal region of the L protein from lymphocytic choriomeningitis virus (LCMV, which is in the same Arenaviridae family as LASV, have identified an endonuclease domain that presumably cleaves the cap structures of host mRNAs in order to initiate viral transcription. Here we present a high-resolution crystal structure of the N-terminal 173-aa region of the LASV L protein (LASV L173 in complex with magnesium ions at 1.72 Å. The structure is highly homologous to other known viral endonucleases of arena- (LCMV NL1, orthomyxo- (influenza virus PA, and bunyaviruses (La Crosse virus NL1. Although the catalytic residues (D89, E102 and K122 are highly conserved among the known viral endonucleases, LASV L endonuclease structure shows some notable differences. Our data collected from in vitro endonuclease assays and a reporter-based LASV minigenome transcriptional assay in mammalian cells confirm structural prediction of LASV L173 as an active endonuclease. The high-resolution structure of the LASV L endonuclease domain in complex with magnesium ions should aid the development of antivirals against lethal Lassa hemorrhagic fever.

  19. Suspected severe fever with thrombocytopenia syndrome in Tongling area:Clinical features in 24 cases%铜陵地区疑似发热伴血小板减少综合征24例临床分析

    Institute of Scientific and Technical Information of China (English)

    沈智勇; 林建; 宋有良

    2014-01-01

    Objective:To analyze the clinical features in patients with suspected severe fever with thrombocytopenia syndrome ( SFTS) prevalent in Tongling area,Anhui,China.Methods:Clinical data were obtained in 24 cases with suspected SFTS undergone diagnosis and treatment in our hospital in the past three years,and reviewed regarding the important clinical manifestations and laboratory findings .Results:Although only 2 were confirmed victims of novel bunyavirus,yet the total 24 cases were commonly characterized by high fever,chill,enlarged superficial lymph nodes,nausea and vomiting,diarrhea,mus-cular soreness,dark stools,haziness,abdominal pain,headache,drowsiness,skin petechiae and ecchymosis,hypotension,coma and shock.Thrombocytope-nia occurred in 87.5%and leucopenia in 75%of the total 24 cases by laboratory detection,who simultaneously had significantly elevated level of aspartate aminotransferase(AST) and alanine transarninase(ALT).Conclusion:Attention should be given in diagnosis of the SFTS due to its absence of notable clinical manifestations in the early stage to prevent misdiagnosis .%目的:分析铜陵地区疑似发热伴血小板减少综合征病例临床特点。方法:回顾收集我院近3年收治的疑似发热伴血小板减少综合征患者临床资料,统计分析所有患者的主要临床表现及相关实验室检查结果。结果:24例疑似SFTS病例中,确诊人感染新型布尼亚病毒病2例;所有疑似病例均有发热,部分病例有畏寒、浅表淋巴结肿大、恶心呕吐、腹泻及肌肉酸痛、黑便、神志淡漠、腹痛、头痛、嗜睡及皮肤瘀点瘀斑、低血压/休克及昏迷等;87.5%的病例有血小板计数减少,75%的病例有白细胞计数减少;24例患者均有肝脏血清生化学指标异常,其中天门冬氨酸转氨酶( AST)水平较丙氨酸转氨酶( ALT)水平升高明显。结论:SFTS病例早期无显著特征性临床表现,易误诊,临床医生需提高认识。

  20. Emerging animal viruses: real threats or simple bystanders?

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    Eduardo Furtado Flores

    2013-10-01

    Full Text Available The list of animal viruses has been frequently added of new members raising permanent concerns to virologists and veterinarians. The pathogenic potential and association with disease have been clearly demonstrated for some, but not for all of these emerging viruses. This review describes recent discoveries of animal viruses and their potential relevance for veterinary practice. Dogs were considered refractory to influenza viruses until 2004, when an influenza A virus subtype H3N8 was transmitted from horses and produced severe respiratory disease in racing greyhounds in Florida/USA. The novel virus, named canine influenza virus (CIV, is considered now a separate virus lineage and has spread among urban canine population in the USA. A new pestivirus (Flaviviridae, tentatively called HoBi-like pestivirus, was identified in 2004 in commercial fetal bovine serum from Brazil. Hobi-like viruses are genetically and antigenically related to bovine viral diarrhea virus (BVDV and induce similar clinical manifestations. These novel viruses seem to be widespread in Brazilian herds and have also been detected in Southeast Asia and Europe. In 2011, a novel mosquito-borne orthobunyavirus, named Schmallenberg virus (SBV, was associated with fever, drop in milk production, abortion and newborn malformation in cattle and sheep in Germany. Subsequently, the virus disseminated over several European countries and currently represents a real treat for animal health. The origin of SBV is still a matter of debate but it may be a reassortant from previous known bunyaviruses Shamonda and Satuperi. Hepatitis E virus (HEV, family Hepeviridae is a long known agent of human acute hepatitis and in 1997 was first identified in pigs. Current data indicates that swine HEV is spread worldwide, mainly associated with subclinical infection. Two of the four HEV genotypes are zoonotic and may be transmitted between swine and human by contaminated water and undercooked pork meat. The

  1. 中国首次分离的Tahyna病毒毒株的生物学特性及分子进化特征分析%Biological and Phylogenetic Analysis of First Isolate of Tahyna Virus in China

    Institute of Scientific and Technical Information of China (English)

    吕志; 付士红; 吕新军; 张松; 童苏祥; 高春代; 姚新华; 梁国栋

    2011-01-01

    2006年夏季,本课题组在新疆喀什地区伽师县库蚊标本中分离到我国首株Tahyna病毒XJ0625株,并发现当地不明原因发热患者存在该病毒感染.本研究通过细胞培养、动物实验、电镜观察、间接免疫荧光及交叉保护中和试验等研究对XJ0625病毒株的细胞易感性、动物致病性、形态学及抗原性等特征进行观察,并应用分子生物学软件对其分子进化特征加以分析.结果发现该毒株可以引起BHK-21细胞病变,乳鼠颅内接种该毒株可以引起死亡.与其他布尼亚病毒形态相似,Tahyna病毒为球形有包膜病毒.该病毒与国际流行Tahayna病毒Bardos92株的抗体作用,在间接免疫荧光试验中呈现阳性荧光信号.空斑减少中和试验结果显示该抗体对XJ0625病毒株的中和效价为1:3 200.核苷酸序列分析显示,该病毒与Bardos92株处于同一个进化分支,二者S节段同源性为91.8%,M节段同源性为81.9%.%In 2006, the first Chinese Tahyna virus isolate (XJ0625) was obtained in Xinjiang province and human infection were found in the same region. In this study, cell culture, animal experiments, electron microscopy, immunofluorescence assay and cross neutralization tests were performed to see the cell susceptibility, animal pathogenicity, morphology and antigenic and other biological characteristics of XJ0625. In addition, molecular biology software was used to analyze the characteristics of molecular evolution. The results showed that BHK-21 cell line was susceptible to XJ0625 and the virus was lethal to suckling mice when injected by intracranial ways. Similar to the other Bunyavirus, Tahyna virus is spherical enveloped virus under electron microscopy. XJ0625 infected cells showed strong fluorescent signal and could be neutralied by immune asities fluid with immnity to protype Tahyna virus Bardos 92. The sequence of the S and M segments showed 91.8% and 81.9% homology with Bardos 92.

  2. Temporal and geographic evidence for evolution of Sin Nombre virus using molecular analyses of viral RNA from Colorado, New Mexico and Montana

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    Calisher Charles H

    2009-07-01

    Full Text Available Abstract Background All viruses in the family Bunyaviridae possess a tripartite genome, consisting of a small, a medium, and a large RNA segment. Bunyaviruses therefore possess considerable evolutionary potential, attributable to both intramolecular changes and to genome segment reassortment. Hantaviruses (family Bunyaviridae, genus Hantavirus are known to cause human hemorrhagic fever with renal syndrome or hantavirus pulmonary syndrome. The primary reservoir host of Sin Nombre virus is the deer mouse (Peromyscus maniculatus, which is widely distributed in North America. We investigated the prevalence of intramolecular changes and of genomic reassortment among Sin Nombre viruses detected in deer mice in three western states. Methods Portions of the Sin Nombre virus small (S and medium (M RNA segments were amplified by RT-PCR from kidney, lung, liver and spleen of seropositive peromyscine rodents, principally deer mice, collected in Colorado, New Mexico and Montana from 1995 to 2007. Both a 142 nucleotide (nt amplicon of the M segment, encoding a portion of the G2 transmembrane glycoprotein, and a 751 nt amplicon of the S segment, encoding part of the nucleocapsid protein, were cloned and sequenced from 19 deer mice and from one brush mouse (P. boylii, S RNA but not M RNA from one deer mouse, and M RNA but not S RNA from another deer mouse. Results Two of 20 viruses were found to be reassortants. Within virus sequences from different rodents, the average rate of synonymous substitutions among all pair-wise comparisons (πs was 0.378 in the M segment and 0.312 in the S segment sequences. The replacement substitution rate (πa was 7.0 × 10-4 in the M segment and 17.3 × 10-4 in the S segment sequences. The low πa relative to πs suggests strong purifying selection and this was confirmed by a Fu and Li analysis. The absolute rate of molecular evolution of the M segment was 6.76 × 10-3 substitutions/site/year. The absolute age of the M segment