Classification of the orbiviruses. Confusion in the use of terms bluetongue virus, bluetongue-like virus, bluetongue-related virus and the overall nomenclature.
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Epidemiology of bluetongue viruses in the American tropics. Regional Bluetongue Team.
A study of the epidemiology of bluetongue viruses is in progress with the collaboration of 11 Central American and Caribbean countries. To date, over 200 bluetongue virus isolates have been obtained from cattle and sheep in sentinel groups distributed in the participating countries. Bluetongue serotypes identified include 1, 3, 6, and 12, virus types not previously recorded in the Western Hemisphere. Although the clinical impact of bluetongue virus infections in this hyperendemic environment appears to be minimal, the ubiquity of infection causes restrictions on the export of ruminant livestock and germ plasm. The stability of the Caribbean region ecosystem and the long-range implications of the interface with the northern temperate bluetongue virus ecosystem are reviewed.
Animals↗
Isolation and identification of a variant of bluetongue virus serotype 11 from a ram in a bluetongue outbreak in western Texas.
A field strain of bluetongue virus was isolated from a blood sample of a ram during an outbreak of bluetongue in November 1985 in western Texas. In this bluetongue outbreak at least 25 of the 2,000 sheep were infected. Isolation was made by intravenous inoculation of 11-day-old embryonated chicken eggs. The serotype was identified as serotype 11 by serum neutralization tests. The genomic pattern on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) of the new isolate is similar to that of bluetongue virus prototype 11. Comparisons were also made with proteins labeled in vivo with [3H]leucine and separated by SDS-PAGE. We conclude that this virus belongs to serotype 11, with slight differences in both genome and protein electrophoretic patterns.
Animals↗
Bluetongue virus isolations from vectors and ruminants in Central America and the Caribbean. Interamerican Bluetongue Team.
A regional prospective study of the epidemiology of bluetongue virus (BTV) serotypes covering 11 countries in Central America and the Caribbean took place between 1987 and 1992. Active surveillance revealed BTV infection to be endemic in the absence of confirmed indigenous cases of bluetongue. During the 6-year span of the study, over 300 BTV isolations were obtained from cattle and sheep. Results of the earlier years of the study were summarized, and surveillance activities in the concluding months of the study from November 1990 to February 1992 were evaluated. Forty-five BTV isolations were made during this time, 44 from sentinel cattle and 1 from a ram with clinical signs compatible with contagious ecthyma. Virus isolation from potential vectors also was attempted, yielding a further 9 BTV isolates from parous Culicoides insignis and C pusillus, 2 BTV isolates from blood-engorged C filarifer, and 1 epizootic hemorrhagic disease virus type-2 isolate from parous C pusillus. Our extensive network of sentinel herds in the region detected BTV-1 as the predominant serotype in Central America in 1991, after an apparent absence of 1 year in the sentinel animals. Other serotypes in Central America at that time included BTV-3 and BTV-6. In Puerto Rico and the Dominican Republic, BTV-4 became the predominant serotype, without detection of BTV-8 and BTV-17, which were common in recent years of the study. The serotypes found in the Caribbean Basin continued to have marked differences from those in North America. The importance of viewing bluetongue as an infection, the distribution of which is determined principally by ecologic factors, is emphasized.
Animals↗
Electrophoretic comparison of the genomes of North American bluetongue viruses, one Australian bluetongue virus, and three other related orbiviruses.
The genomes of U.S. bluetongue viruses, an Australian bluetongue virus, and three other related orbiviruses were analyzed by polyacrylamide gel electrophoresis. The genomes were comprised of ten segments of double-stranded (ds) RNA. Estimates of the molecular weights of the dsRNA segments revealed that the U.S. bluetongue serotypes were remarkably similar. Although the dsRNA profiles of the viruses exhibited common segments, each virus had a distinct dsRNA profile. The usefulness of the genome analysis as a diagnostic tool for identification and for epidemiologic studies is discussed.
Arboviruses↗
Comparison of bluetongue type 20 with certain viruses of the bluetongue and Eubenangee serological groups of orbiviruses.
The genome of bluetongue virus type 20 consists of 10 segments of double-stranded RNA each of which contains unique sequences as determined by oligonucleotide mapping. The 10 polypeptide products of the virus genome were detected in virus-infected cells, and in pulse--chase experiments there was no secondary cleavage of the primary gene products. Using stringent conditions for RNA--RNA reassociation, no significant homology could be detected between the genomes of bluetongue type 20 isolated in Australia and representative serotypes isolated in other geographic regions. The results suggest sequence divergence between geographically isolated viruses and not the recent introduction of a bluetongue virus into Australia.
Base Sequence↗
Bluetongue in cattle: effects of vector-transmitted bluetongue virus on calves previously infected in utero.
Three of 7 principal calves, after a challenge of immunity exposure by bites of bluetongue (BT) virus-infected Culicoides variipennis, became latently infected with BT virus. These calves were born to heifers infected with the homologous virus by bites of C variipennis at 60 or 120 days' gestation. Latent BT virus infection was detected by isolation of BT virus from washed erythrocyte samples obtained from the calves at 57, 100 to 102, 200 to 202, 300 to 302, and 400 to 402 days after challenge of immunity and from 1 of the calves over 5 years after challenge of immunity. The 3 latently infected calves were healthy; 2 were immunologically competent and 1 was immunologically incompetent to develop detectable BT virus antibodies in their blood. Bluetongue virus infection was detected (by viral isolation) in 2 other principal calves during the challenge of immunity, but they were not considered latently infected. The latter 2 calves were immunologically incompetent to develop detectable BT virus antibodies.
Animals↗
Comparative susceptibility of a canine cell line and bluetongue virus susceptible cell lines to a bluetongue virus isolate pathogenic for dogs.
Recently, bluetongue virus (BLU) serotype 11 was detected in diseased dogs that had been inoculated with live attenuated vaccine contaminated with this serotype of bluetongue virus (Akita et al., 1994). For various laboratory tests, BLU can be propagated in different cell cultures. No information was found in the literature about the possibility of propagating this virus in canine cells. To determine whether the BLU isolate from the contaminated canine vaccine (BLU-vac) is unique in its ability to replicate in canine cells, this virus was studied in parallel with U.S. prototype strains of BLU (serotypes 2, 10, 11, 13, and 17), in hamster lung (HmLu-1) and canine kidney (MDCK) cell cultures. In HmLu-1 cell cultures, the BLU-vac produced cytopathic effect (CPE) of the same type as the U.S. prototype BLU strains by 4 to 6 d postinoculation. In MDCK cell cultures, all of the BLU strains tested were able to replicate but did not produce CPE. The BLU-inoculated MDCK cells became persistently infected, and these cultures continued to produce infectious BLU even after six serial passages over 2 1/2 mo. In none of these cultures was CPE observed. In mixed cultures containing both HmLu-1 and MDCK cells, CPE first affected the HmLu-1 islands; subsequently, CPE spread also to the areas with MDCK cells. The silent persistent infection of the MDCK cells with BLU indicates that more stringent screening of the cells used in the production of live vaccines for various contaminating viruses is necessary.
Animals↗
Epidemiologic study of bluetongue viruses in Central America and the Caribbean: 1986-1988. Regional Bluetongue Team.
Results of a prospective serologic and virologic study of ruminant livestock in Central America and the Caribbean islands revealed bluetongue virus (BTV) to be enzootic in the 9 countries participating in the study. Bluetongue virus serotypes 1, 3, 6, and 12 were isolated from sentinel animals. To the authors' knowledge, these are the first isolations of BTV from the region studied and the first isolations of these serotypes in the Western Hemisphere. Clinical disease attributable to BTV infection was not observed in sentinel animals. The incidence pattern, with respect to age and geographic location, was determined. The need to evaluate the epizootiologic features or arthropod-borne viruses (arboviruses) on a regional ecologic basis is stressed.
Animals↗
Multiple bluetongue virus-cloned genetic probes: application to diagnostics and bluetongue virus genetic relationships.
A shotgun-cloning method incorporating all 10 bluetongue virus genome segments can simultaneously produce complete and partial copies of any of the genome segments. We report here 4 different cloned probes derived from 3 genome segments and individually defined by different hybridization recognition capabilities. One probe hybridized strongly with all 5 United States prototype strains of the 5 different bluetongue virus (BTV) serotypes existing in the United States and, as such, is a strong candidate for a broad BTV diagnostic probe in the United States. Another probe derived from genome segment 2 of BTV-17 hybridized only with the BTV-17 prototypic serotype, thereby demonstrating serospecific hybridization diagnostic potential. The implications for diagnostic and genetic relationship studies on BTV, using various genetic probes, are discussed.
Animals↗
Expression of the major core structural protein (VP7) of bluetongue virus, by a recombinant capripox virus, provides partial protection of sheep against a virulent heterotypic bluetongue virus challenge.
A recombinant capripox virus was constructed containing a cDNA copy of genome segment 7 of bluetongue virus (BTV) serotype 1 from South Africa (BTV 1SA), which expressed high levels of the major BTV core protein VP7 in infected lamb testis (LT) cells. Sheep vaccinated with this recombinant virus developed antibodies to VP7 (detected by ELISA) but no neutralizing antibodies to either the homologous or heterologous BTV serotype, prior to challenge (BTV 1 or BTV 3, respectively). Following challenge with a virulent heterotypic strain of BTV (BTV3 SA), all of the animals developed clinical signs of disease, indicating that they were infected and that the challenge virus did replicate. While all of the control animals died, six of the eight animals that were vaccinated with the recombinant capripox virus expressing VP7 recovered fully. This is the first report of a significant level of cross serotype protection against the lethal effects of a challenge with virulent BTV, produced by vaccination with a single BTV core protein, which did not generate a neutralizing antibody response.
Animals↗
Dot immunobinding assay for the detection of bluetongue virus antibodies in sheep experimentally inoculated with bluetongue virus type 1.
Dot immunobinding assay (DIA) was evaluated for the detection of bluetongue virus (BTV) antibodies in sheep experimentally inoculated with BTV 1. Serum samples collected on 14, 21, 28, 43 and 60 day post infection (dpi) were positive for precipitating antibodies by the agar gel precipitation test (AGPT) while antibodies could be detected as early as 7 dpi by DIA and ELISA. Virus neutralizing antibodies were detected first at 14 dpi. The sensitivity of the four tests was compared on the same serum samples collected at different intervals. The results indicated that DIA was more sensitive than AGPT and the serum neutralization test and as sensitive as ELISA. Thus due to sensitivity simplicity and economy, DIA could replace AGPT for diagnosis and serological survey for BTV infection in animals.
Animals↗
Further studies on bluetongue and bluetongue-related orbiviruses in the Sudan.
The seasonal incidence of bluetongue virus (BTV) in Central Sudan is related primarily to fluctuations in the prevalence of the vector, Culicoides imicola. Population densities of this midge begin to rise with the onset of precipitation and peak during October, before falling sharply at the end of the rainy season in November. These are also the months of BTV transmission. Populations of C. schultzei, the commonest midge in Central Sudan, are also related to the rainy season but this species does not seem to be involved with BTV transmission. BTV serotype 2 was isolated from C. imicola confirming the status of this midge as a known vector but a second isolate of the same serotype was made from a mixed pool of Culicoides not including C. imicola. This suggests that BTV transmission in the Sudan may involve more than one species of Culicoides. Epizootic haemorrhagic disease virus (EHDV) serotype 4 and a palyam virus were isolated from C. schultzei which indicates that this species may be involved in the transmission of BT-related viruses. Seven further virus isolates from sentinel calves at Shambat (Khartoum) confirmed the presence of BTV serotypes 1, 4 and 16, and an untyped EHDV (designated 318) in the Sudan. All of the viruses isolated and identified during the course of this work are recorded from the Sudan for the first time.
Animals↗
Epidemiology of bluetongue in Central America and the Caribbean: initial entomological findings. Regional Bluetongue Team.
Forty-four species of Culicoides (Diptera: Ceratopogonidae) were caught in insect light traps during the first 2 years of studies on the epidemiology of bluetongue virus in the Caribbean and Central America. Traps were operated near sentinel ruminants which were bled monthly for serologic evaluation and then virus isolation. More than 570,000 individuals were identified. Culicoides insignis Lutz accounted for 90% of the catch, C. filarifer Hoffman/C. ocumarensis Ortiz 5%, C. furens Poey 3% and C. pusillus Lutz 2%. Other species accounted for less than 1% of the total catch. Sentinel ruminants became seropositive when C. insignis populations were high at many study sites. At a few sites C. pusillus and C. filarifer/C. ocumarensis were predominant or were present in large numbers during seroconversions of sentinels. Virus isolations were obtained from sentinel ruminants during times when these same species were present in large populations.
Animals↗
Seasonal transmission of bluetongue virus by Culicoides sonorensis (Diptera: Ceratopogonidae) at a southern California dairy and evaluation of vectorial capacity as a predictor of bluetongue virus transmission.
Vectorial capacity of Culicoides sonorensis Wirth & Jones for the transmission of bluetongue (BLU) virus was examined at a southern California dairy from January 1995 to December 1997. Insects were collected one to two times per week in five CDC-type suction traps (without light) baited with CO2 at a constant release rate of 1,000 ml/min. BLU virus was detected in midges collected from May through December with an estimated overall infection rate of 0.08%. The BLU virus infection rate of field-captured midges was not correlated with sentinel calf seroconversions to BLU virus. Sentinel calf seroconversions were highly seasonal, occurring from August through November with most calves seroconverting during September and October. Vector competence of field-collected nulliparous flies fed a locally acquired serotype of BLU virus in the laboratory was stable among years (17-23%). Vectorial capacity was strongly correlated with BLU virus transmission (measured by sentinel calf seroconversions) during 1995 and 1996, but not during 1997. Host biting rate estimated for traps nearest to the sentinel calves was the index best correlated with BLU virus transmission for all study years and was most highly correlated with sentinel seroconversions 4 wk later. The utility of vectorial capacity and its component variables is discussed for this system.
Animals↗
Bluetongue in cattle: repeated exposure of two immunologically tolerant calves to bluetongue virus by vector bites.
A heifer and steer that were immunologically tolerant to bluetongue (BT) virus became immunologically competent after repeated exposures by bites of BT virus-infected Culicoides variipennis. Immunologic tolerance ended in the heifer at 25 months of age, after the 2nd exposure to the virus, and in the steer at 22 months, after the 4th exposure. High hemic concentrations of BT virus were detected in both animals after they became immunologically competent, but neither developed an overt BT clinical response. The steer died suddenly and extensive pathologic changes were observed at necropsy.
Animals↗
A bluetongue serogroup-reactive epitope in the amino terminal half of the major core protein VP7 is accessible on the surface of bluetongue virus particles.
Immunoelectron microscopy has been used to confirm that the core protein VP7 is accessible on the surface of bluetongue virus (BTV) particles. Monospecific antibodies generated to vaccinia virus-expressed VP7 and an anti-VP7 monoclonal antibody (MAb 20E9) bound to native virus particles and were localized by protein A-gold. In contrast, MAb 20E9 labeled directly with gold failed to gain access and bind, suggesting that VP7 is neither adventitiously adsorbed to the virion surface nor exposed in a manner such as protrusion through the outer capsid. Thus the surface layer of BTV may be considered as a net which only partially obscures the underlying core particle. Sequencing of VP7 revealed it to be an extremely hydrophobic protein, 350 amino acids in length with cysteine residues at positions 15, 65, and 154. Examination of VP7 in the cytosol of cells infected with either BTV or a vaccinia virus recombinant expressing VP7 indicated that the protein may exist as an oligomer, whose constituent monomers are not linked by intermolecular disulfide bonds. The cysteine residues in sodium dodecyl sulfate (SDS)-denatured, dithiothreitol (DTT)-treated VP7 were labeled with the fluorescent iodoacetamide AEDANS and the protein was cleaved by V8 protease. The size of the labeled peptides and knowledge of the location of potential V8 cleavage sites suggested that the enzyme cleaved VP7 at three locations (glutamic acid residues at positions 61, 104 (or 108), and 132 (or 134 or 135). Analysis of the fluorescent peptides generated by V8 protease cleavage of VP7 labeled with AEDANS in the absence of DTT (i.e., with any putative intramolecular disulfide bonds intact) suggested that the cysteine at position 154 was the only one accessible to AEDANS. The cysteines at positions 15 and 65 may therefore be linked via a disulfide bond. Denaturation of VP7 with SDS did not eliminate the capacity of the protein to bind MAb 20E9. However, the sensitivity of the epitope to reduction and acetylation and its resistance to either of these processes alone suggest that it may be located near a disulfide bond linking cysteines at positions 15 and 65. Confirmation that the epitope lay in the amino-terminal half of the VP7 came from immunoelectron microscopy experiments in which thin sections of bacteria expressing the complete VP7 and the amino-terminal half were probed with MAb 20E9 and protein A-gold.
Amino Acid Sequence↗
Analysis of genome segments from six different isolates of bluetongue virus using RNA-RNA hybridisation: a generalised coding assignment for bluetongue viruses.
Nucleic acid probes prepared directly from bluetongue virus (BTV) genomic double-stranded RNA (dsRNA) have been used to identify the functionally equivalent genome segments from six distinct isolates of BTV after their separation in both agarose and polyacrylamide gel electrophoresis systems. Variations in the rate, and in one case the order, of migration of the equivalent genome segments from different viruses was detected in the polyacrylamide gel system. However, the genomic dsRNA profiles of eleven BTV isolates were found to be identical when analysed by agarose gel electrophoresis. Functionally equivalent genome segments from the six viruses that were analysed were found to migrate in identical relative positions in this gel system. From these data we propose a modified version of the protein coding assignments published for BTV 1 South Africa (Mertens et al., 1984) in which the identification of the genome segments would be based upon their order of migration in the agarose rather than the polyacrylamide gel system. The modified coding assignments, unlike the original assignments, would be applicable to all of those viruses analysed and appear likely to be valid for all normal BTV isolates.
Bluetongue virus↗