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At least 19 recordsLinked to original sources

PubMed1990

RNA probes detect nucleotide sequence homology between members of two different nairovirus serogroups.

Cloned cDNA derived from the small (S) and medium (M) genomic RNA segments of Dugbe (DUG) virus, isolate ArD44313, a member of the Nairobi sheep disease (NSD) serogroup of nairoviruses (family, Bunyaviridae) was used to prepare 32P-labelled DNA and RNA probes. The S and M segments of six isolates of DUG virus all hybridised to both DNA and RNA probes, although the M segment of isolate KT281/75 reacted only weakly. Of nine other nairoviruses tested, representing all the six other serogroups within the Nairovirus genus, none hybridised to the DNA probes. However, under conditions of low stringency, the DUG S and M RNA probes hybridised to the respective S and M segments of Ganjam (GAN) virus (another member of the NSD serogroup). The DUG S RNA probe also hybridised to the S segments of Crimean-Congo haemorrhagic fever (CCHF) virus and Hazara (HAZ) virus (members of the CCHF serogroup). The indicated sequence relationships between DUG, GAN, CCHF and HAZ viruses show that the NSD serogroup is more closely related to members of the CCHF serogroup than it is to nairoviruses of the other five serogroups.

Animals↗

PubMed1992

Expression of the nucleocapsid protein of Dugbe virus and antigenic cross-reactions with other nairoviruses.

The small (S) RNA segment of Dugbe (DUG) virus (Nairovirus, Bunyaviridae) encodes a single protein, the nucleocapsid (N) protein, of M(r) 49.4 kDa. cDNA derived from the complete coding region for the N protein was cloned into Autographa californica nuclear polyhedrosis virus (AcNPV) under control of the polyhedrin promoter and used to infect Spodoptera frugiperda insect cells. Western blotting analysis using monoclonal antibodies demonstrated the production of DUG N protein in the infected cells. Monoclonal and polyclonal antibodies to the N protein of Crimean-Congo haemorrhagic fever (CCHF) virus were found to cross-react weakly with the baculovirus expressed DUG N protein by Western blotting. When used in an enzyme linked immunoassay (ELISA), the DUG N protein reacted with polyclonal mouse immune ascitic fluids raised against either CCHF or Hazara viruses (both members of the CCHF serogroup of nairoviruses). Cross-reactions between DUG virus (Nairobi sheep disease serogroup) and members of other nairovirus serogroups were not detected.

Antigens, Viral↗

PubMedDate not supplied

Infection and coding strategies of arenaviruses, phleboviruses, and nairoviruses.

The infection and coding strategies of three groups of negative-stranded RNA viruses (arena viruses, phleboviruses, and nairoviruses) that include the etiologic agents of hemorrhagic disease in humans have been studied. Arenaviruses have two viral RNA species. The smaller RNA species (S) codes for the viral nucleoprotein (N protein) and for the viral glycoprotein species (G1 and G2, which are derived from a precursor glycoprotein, GPC). The S RNA has an ambisense arrangement. The proteins are translated from subgenomic mRNA species (viz., N protein from a viral-complementary mRNA and glycoprotein from a viral-sense mRNA). The larger arenavirus RNA species (L) is presumed to code for the viral transcriptase/replicase. Phleboviruses and nairoviruses are members of the Bunyaviridae. They both have three species of viral RNA. Other than the sizes of the viral proteins and the viral RNA species, virtually nothing is known about the coding strategy of nairoviruses. Phleboviruses have an ambisense coding arrangement to their smallest (S) RNA species. This S RNA codes for the viral N protein (translated from a viral-complementary mRNA) and a nonstructural protein (translated from a viral-sense mRNA). The middle-size (M) RNA of phleboviruses codes for a precursor to the viral glycoproteins (translated from a viral-complementary mRNA). The largest viral RNA (L) is presumed to code for the viral transcriptase/replicase.

Animals↗

PubMed1990

Studies on the pathogenicity of a nairovirus, Dugbe virus, in normal and immunosuppressed mice.

Susceptibility to lethal infection with the KT281/75 strain of the tick-borne nairovirus, Dugbe virus, was similar in an outbred strain and several inbred strains of mice. For the outbred strain, both neural and extraneural routes of virus inoculation resulted in lethal infection, but susceptibility decreased with age and only intracerebral inoculation produced a lethal infection in adults. In newborn mice, subcutaneous (s.c.) inoculation of virus (analogous to a tick-bite) produced a disseminated infection, titres being highest in the upper respiratory tract (URT), spleen and liver at 5 days post-inoculation (p.i.), the heart at 7 days p.i. and brain by 8 days p.i. In neonates inoculated intranasally (i.n.), by contrast, virus spread rapidly from the URT to the brain by 2 days p.i., in the absence of a detectable viraemia. Virus was undetectable in the blood of s.c. and i.n. inoculated adults; in the former, virus replication was limited to the site of inoculation, and in the latter virus grew in the respiratory tract and again spread to the brain. Immunosuppression of i.n. inoculated adult mice with cyclophosphamide produced some mortality indicating that host defences are important in protecting the adult, especially as newborn and adult lung tissue were equally able to support the growth of Dugbe virus in culture. The similarity between the pattern of Dugbe virus infection in the mouse and that of other, more pathogenic nairoviruses suggests that, although haemorrhagic disease was not observed, this may be a useful model for studying the genetic basis of nairovirus virulence and for testing vaccines and anti-viral drugs.

Animals↗

PubMed1991

Structure and morphogenesis of Dugbe virus (Bunyaviridae, Nairovirus) studied by immunogold electron microscopy of ultrathin cryosections.

We have studied the structure and morphogenesis of Dugbe (DUG) virus (Bunyaviridae, Nairovirus) in cultured porcine kidney (PS) cells and a tick cell line (Ra 243) using immunogold electron microscopy. DUG virus is a tickborne arbovirus, considered to be a low health hazard, that is antigenically and genetically related to Crimean Congo haemorrhagic fever (CCHF) virus (Marriott et al., 1990). We have investigated the maturation and intracellular transport of DUG virus particles as a model for other more pathogenic nairoviruses using monoclonal antibodies for immunogold labelling of ultrathin cryosections and immunofluorescence techniques. The spherical DUG virus particle measures about 90 nm in diameter, with a 5 nm thick membrane covered by 5-7 nm long projections or "spikes". These projections form hollow cylindrical morphological units, about 5 nm in diameter. DUG virus infection caused only a slight cytopathogenic effect in mammalian cells and none in tick cells. DUG virus particles assembled by budding from the Golgi complex, where the DUG virus glycoprotein G1 accumulated in vesicles originating from Golgi cisternae. The nucleocapsid protein N accumulated in scattered foci throughout the cytoplasm, and this appears to be related to the limited maturation of DUG virus particles that occurred. The reduced number of budding virus particles observed in tick cells was correlated with the reduced cytopathology observed.

Animals↗

PubMed1992

Dugbe Nairovirus M RNA: nucleotide sequence and coding strategy.

The coding assignments of the medium-sized (M) RNA segment of the Dugbe (DUG) virus (Nairovirus, Bunyaviridae) were investigated. The complete nucleotide sequence of 4888 nucleotides (nt) contained one long open reading frame in the viral complementary RNA, extending from an AUG start codon at nt 48-50 to a stop codon at nt 4701-4703 (numbered from the 5' terminus of vcRNA). Comparison of the terminal sequences with the ends of the DUG S segment revealed sequence identity between the first nine nucleotides of both segments. No sequence homologies were found with the M segments of other members of the Bunyaviridae, or with their polypeptide products. Expression of portions of the DUG M open reading frame in Escherichia coli demonstrated the carboxyl terminal region of the M open reading frame codes for the G1 structural glycoprotein, which is the target for neutralising antibodies. Confirmation of this assignment was obtained by sequencing the amino terminus of the G1 protein. Two nonstructural glycoproteins which share epitopes with G1 were identified in virus-infected cells, one of which (85 kDa) is processed over a period of several hours to produce G1. The G2 coding region was located upstream of the G1 sequence. The region between the carboxyl terminus of G2 and the 5' end of the long open reading frame apparently encodes a nonstructural protein of about 70 kDa, which is a precursor of the G2 protein.

Amino Acid Sequence↗

PubMed1992

Mechanisms of neutralization of a nairovirus (Dugbe virus) by polyclonal IgG and IgM.

Dugbe virus is a member of the nairovirus genus of the Bunyaviridae. Purified polyclonal anti-Dugbe virus IgG, which neutralized greater than 99.5% of virus, reduced attachment of virus to BSC-1 cell monolayers by only 36%. A 100-fold lower concentration neutralized virus by 88%, and had no effect upon attachment. Neutralizing IgG did not affect the ability of Dugbe virus to be internalized by or to fuse with BSC-1 cells. This suggests that IgG neutralization occurs largely at a stage subsequent to primary uncoating. Purified polyclonal anti-Dugbe virus IgM neutralized infectivity and had no effect on the attachment of virus to cells, but inhibited internalization of virus by about 50%. Thus IgM neutralizes partly by interfering with entry of virus and partly by a post-entry event. Neutralization by intermediate concentrations of IgM was enhanced 20-fold in the presence of complement. At high concentrations of IgM, complement-dependent neutralization declined. This is probably due to IgM binding in a planar rather than crab conformation, which does not expose the complement binding sites. Aggregation occurred only at relatively low concentrations of immunoglobulin. Electron microscopy and reactivation of infectivity by vortexing suggested that aggregation makes only a minor contribution to neutralization by IgG or IgM.

Animals↗

PubMed1990

Coding strategy of the S RNA segment of Dugbe virus (Nairovirus; Bunyaviridae).

The S RNA segment of Dugbe (DUG) virus (Nairovirus; Bunyaviridae) was sequenced from three overlapping cDNA clones and by primer extension. The S RNA is 1712 nucleotides in length and contains one large open reading frame (ORF) of 1326 nucleotides coding for a 49.4-kDa protein on viral complementary (vc) RNA. This protein in size corresponds to the DUG nucleocapsid (N) protein (P. Cash, 1985, J. Gen. Virol. 66, 141-148). The 49.4-kDa product was expressed as a fusion protein with beta-galactosidase in Escherichia coli cells and confirmed as DUG N protein by Western blotting with DUG N-specific monoclonal antibody. An additional ORF of 150 nucleotides coding for a possible 5.9-kDa protein is present in the +1 reading frame, 3' to the N protein ORF on vcRNA. DUG S segment mRNA was found to be essentially full length. No evidence was obtained for the existence of a smaller mRNA species that could code for a 5.9-kDa protein. Comparisons of the DUG S RNA sequence and predicted N protein amino acid sequence, with the respective sequences of snowshoe hare, La Crosse (bunyaviruses), Punta Toro, Sandfly fever Sicilian (phleboviruses), and Hantaan (hantavirus) viruses, failed to detect any sequence similarity, although the genomic structure of DUG S RNA is similar to that of the S RNA segment of Hantaan (HTN) virus.

Amino Acid Sequence↗

PubMed1985

Polypeptide synthesis of Dugbe virus, a member of the Nairovirus genus of the Bunyaviridae.

The replication of Dugbe (DUG) virus, a member of the Nairovirus genus of the Bunyaviridae, has been investigated. During the infection of BS-C-1 cells a virus-specific c.p.e. was initially observed followed by recovery of the cell monolayer but with continued production of infectious virus. Six DUG virus-induced polypeptides were identified with apparent molecular weights, determined by gel electrophoresis, of 92000 (p92), 82000 (p82), 77000 (p77), 52000 (p52), 48000 (p48) and 34000 (p34). The polypeptides p77 and p34 were detected in purified DUG virions but not in extracts of virus-infected cells pulse-labelled with [3H]leucine. Polypeptides p48 and p52 were found in both purified virus preparations and in extracts of infected cells. p82 and p92 were found only in lysates of infected cells. When two-dimensional gel electrophoresis was used to analyse infected cells, p48 was found to have a net positive charge.

Animals↗

PubMed1985

The proteins and RNAs specified by Clo Mor virus, a Scottish Nairovirus.

The proteins and RNAs of Clo Mor virus have been analysed. Virus-specific proteins in infected cells had previously been identified by isotopic labelling and radioimmunoprecipitation; these were three glycoproteins (mol. wt. 115K, 90K and 80K) and an unglycosylated nucleocapsid (N) protein (50K). We have performed pulse-chase experiments which indicated that the 115K protein is processed to give the 90K and 80K proteins, while a 45K protein was detected in released virions after prolonged chase. Translation in vitro of mRNA extracted from Clo Mor virus-infected cells resolved only the N protein. Three species of RNA were extracted from Clo Mor virus intracellular nucleocapsids and have been designated L (11000 to 13000 bases), M (6300 bases) and S (1900 bases). The processing of viral proteins and the sizes of RNAs are characteristic of the Nairovirus genus of the family Bunyaviridae.

Bunyaviridae↗

PubMed1987

Field and laboratory investigation of Crimean-Congo haemorrhagic fever virus (Nairovirus, family Bunyaviridae) infection in birds.

In November 1984 a case of Crimean-Congo haemorrhagic fever (CCHF) occurred in a worker who became ill after slaughtering ostriches (Struthio camelus) on a farm near Oudtshoorn in the Cape province of South Africa. The diagnosis was confirmed by isolation of CCHF virus from the patient's serum and by demonstration of a specific antibody response. It was suspected that infection was acquired either by contact with ostrich blood or by inadvertently crushing infected Hyalomma ticks while skinning ostriches. Reversed passive haemagglutination-inhibition antibody to CCHF virus was detected in the sera of 22/92 ostriches from farms in Oudtshoorn district, including 6/9 from the farm where the patient worked, but not in the sera of 460 birds of 37 other species. In pathogenicity studies domestic chickens proved refractory to CCHF infection, but viraemia of low intensity (maximum titre 2.5 log10 mouse ic LD50/ml) followed by a transient antibody response occurred in blue-helmeted guinea fowl (Numidia meleagris). These results offer the first direct evidence that some bird species are susceptible to CCHF virus infection.

Animals↗

PubMed1989

Difference in vector competence of two species of sympatric ticks, Amblyomma variegatum and Rhipicephalus appendiculatus, for Dugbe virus (Nairovirus, Bunyaviridae).

Amblyomma variegatum was shown to be a competent vector of Dugbe (DUG) virus whereas Rhipicephalus appendiculatus was not. When DUG virus was taken up orally by A. variegatum nymphs, during capillary feeding, the virus replicated and persisted through moulting to the following adult stage. In contrast, although DUG virus replicated in capillary fed R. appendiculatus nymphs, the virus did not persist trans-stadially into the adult stage. If the oral route of infection was by-passed by direct inoculation into the haemocoel, DUG virus replicated and survived trans-stadially in both tick species, and was subsequently transmitted during feeding. The different responses of R. appendiculatus to oral and intra-coelomic routes suggest that DUG virus is able to infect the gut of this tick species, but that release of the virus from the gut is inhibited.

Animals↗

PubMed1985

Experimental studies on the replication and dissemination of Qalyub virus (Bunyaviridae: Nairovirus) in the putative tick vector, Ornithodoros (Pavlovskyella) erraticus.

A study was undertaken to determine if the argasid tick, Ornithodoros (Pavlovskyella) erraticus, can serve as a biological vector of Qalyub (QYB) virus. The suckling mice used as viremic vertebrate hosts were acceptable hosts for all feeding stages of this tick and developed relatively high titered viremias (4.4-6.5 log10PFU/ml) 24-120 hr post intracerebral inoculation. Larval, nymphal, and adult ticks became orally infected with QYB virus after ingesting 4.4-6.4 log10PFU/ml. The overall infection rate for all experiments was 67/205 and virus was recovered up to day 179 postfeeding. Incubation of known quantities of QYB virus with uninfected triturated tick tissues did not result in any appreciable virus inactivation. QYB viral antigen was detected by immunofluorescence primarily in the tick midgut posterior diverticula cells. First and second instar nymphs orally infected as larvae did not individually transmit QYB virus to suckling mice; however they successfully transmitted the virus when feeding in groups of 11-20 per mouse. Three out of fourteen of the orally-infected male and female ticks individually transmitted QYB virus orally to suckling mice. Organ titrations of ticks orally exposed to QYB virus demonstrated virus primarily in midgut tissues; dissemination to other organ systems was discovered in only 1 tick after 142 days extrinsic incubation. Vertical transmission of virus from infected female ticks to progeny was not demonstrated. Four of the 39 ticks in our colony were infected with a spirochete; presumably, Borrelia crocidurae. O. (P.) erraticus apparently satisfies the conditions that would implicate this species as a biological vector of QYB virus and is the only known arthropod from which this virus has been isolated in nature.

Animals↗

PubMed1989

Experimental infection of six species of ixodid ticks with Dugbe virus (family Bunyaviridae, genus Nairovirus).

The vector potential of each of 6 species of colonized North American and African ixodid ticks was assessed by intracoelomic inoculation with Dugbe virus (IbAr 1792, 14th passage in suckling mouse brain) and viral titers were monitored after selected incubation periods. Persistence of Dugbe virus for greater than or equal to 53 days in 5 species (Dermacentor andersoni, D. variabilis, Amblyomma americanum, Rhipicephalus appendiculatus, and R. sanguineus) indicates that infection occurred. Viral titers were significantly higher in female vs. male D. variabilis, R. appendiculatus, and A. americanum after blood feeding. Blood feeding had no significant effect on the viral titers of either female or male R. sanguineus. D. andersoni males also exhibited no significant change in viral titers after blood-feeding, but 100% (20/20) of drop-off females and 96% (24/25) of post-oviposition females (36 days postinoculation) contained no detectable virus even though virus was still found in unfed specimens less than or equal to 124 days postinoculation. Virus was not recovered from greater than 30,000 1st generation progeny (eggs, larvae, nymphs, adults) collected as eggs from inoculated female D. andersoni, D. variabilis, R. sanguineus, and R. appendiculatus 27-51 days postinoculation. R. sanguineus and R. appendiculatus transmitted Dugbe virus to guinea pigs when allowed to feed 1-3 weeks postinoculation.

Africa↗

PubMed1991

Dissemination, replication, and trans-stadial persistence of Dugbe virus (Nairovirus, Bunyaviridae) in the tick vector Amblyomma variegatum.

The dissemination and replication of Dugbe (DUG) virus and its tissue tropisms in the tick vector Amblyomma variegatum were examined by immunohistochemical analysis using specific antibody, in situ hybridization with a viral-complementary riboprobe, and infectivity assays of dissected tissues. Dugbe virus was localized in both unfed and feeding adults inoculated as nymphs or orally infected by capillary feeding, and in nymphs infected by capillary feeding. In non-feeding ticks, the main sites of DUG virus replication were the epidermis, hemocytes associated with loose connective tissue, and a small number of phagocytic digestive cells in the gut lumen. Virus infectivity in the hemolymph was associated entirely with hemocytes. Dugbe viral antigen or infectivity was not detected in the salivary glands until after the start of feeding. Viral titers in the salivary glands of feeding ticks were about ten-fold higher than in gut, ovary, or loose connective tissue. The level of infection decreased during molting and increased during feeding. Viral particles and pathologic effects were not detected in infected ticks. The primary site of trans-stadial persistence of DUG virus is the hemocytes. Tick hemocytes and other motile cells may be important in the transmission of persistent virus infection from one cell or organ to another by diapedesis.

Animals↗

PubMed1986

Avalon virus, Sakhalin group (Nairovirus, Bunyaviridae) from the seabird tick Ixodes (Ceratixodes) uriae White 1852 in France.

Nine strains of Avalon virus were isolated from Ixodes uriae ticks collected in the Cape Sizun seabird reserve, Brittany, from 1979 to 1985, during a longitudinal study of consequences of tick-borne infections for kittiwakes (Rissa tridactyla). Avalon virus strains isolated in France proved difficult to study owing to the weak infectious titres they exhibited in suckling mice or cultured cells. However, some interesting data concerning the ecology of virus infection and the morphology of the virions were obtained and are discussed.

Animals↗

PubMed1983

Antigenic variants of the Soldado virus (Nairovirus, Bunyaviridae) isolated in different parts of the world.

Antigenic analysis of 9 strains of the Soldado virus, originating from seven different countries of Central and Northern America, Africa, Northern Europe and of the Pacific area was carried out by means of complement fixation (CF) and immunodiffusion (ID) tests. Three strains isolated from Ornithodoros (A.) capensis, including the reference Trinidad strain, were found to be related within the same relatively heterogeneous sub-group C, being characteristic for the New World. Five other strains isolated from Ornithodoros (A.) maritimus were classified to form an another, much more homogeneous subgroup M, characteristic for the Old World (related to the Ireland strain). The South Africa strain, isolated form Ornithodoros (A.), probably capensis, could not be classified in either of the above two subgroups. Thus, the Soldado virus in fact represents a complex of more or less closely related strains, but not a single virus.

Antigens, Viral↗

PubMed1992

Comparison of the S RNA segments and nucleoprotein sequences of Crimean-Congo hemorrhagic fever, Hazara, and Dugbe viruses.

The S RNA segments of the nairoviruses Crimean-Congo hemorrhagic fever (CCHF) virus (Chinese isolate) and Hazara (HAZ) virus were cloned and sequenced from PCR products. The RNAs comprise 1672 and 1677 nucleotides, respectively, and each encodes a protein in the viral complementary strand (54.0 and 54.2 kDa, respectively). The deduced protein sequences show homology to each other and to the nucleoprotein of Dugbe (DUG) nairovirus, although both the CCHF and HAZ viral proteins are larger. Alignment of the nucleoprotein sequences of CCHF, HAZ, and DUG viruses show that the CCHF and HAZ sequences are somewhat more closely related to each other (60.0% identity) than either is to the DUG sequence (55.4 and 53.0% identity, respectively); 39.5% of residues are identical across all three proteins. The carboxyl-terminus of DUG N protein shows a 40-residue deletion relative to the N proteins of the other two viruses.

Amino Acid Sequence↗