Sensitivity of bluetongue virus to lipid solvents, trypsin and pH changes and its serological relationship to arboviruses

Svehag, Sven‐Eric; L, Leendertsen; Gorham, J. R.

doi:10.1017/s0022172400040614

Cited by 21 publications

(8 citation statements)

References 20 publications

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“…Rotavirus infectivity is increased by the presence of trypsin in the culture medium (Babiuk et al, 1977;Almeida et al, 1978;Graham & Estes, 1980), and are also like EHDV, found in extracellular aggregates (Esparza et al, 1980). However, we found that trypsin inactivates EHDV as found by Svehag et al (1966) for bluetongue virus. The large increases in infectivity following sonication monitored by electron microscopy showed the disappearance of aggregates and an increase in intact virus particles.…”

Section: Electron Microscopic Observationssupporting

confidence: 44%

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Replication and Release of Epizootic Haemorrhagic Disease Virus in BHK-21 Cells

Lewanczuk

Yamamoto

1982

Journal of General Virology

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SUMMARYEpizootic haemorrhagic disease virus (EHDV) was seen by light and electron microscopy to replicate in perinuclear locations. Tubules, paracrystals and virus matrices were associated with replication sites. As infection proceeded, aggregates of virus migrated towards the cell periphery, resulting in cell membrane rupture near the virus aggregate with the subsequent release of the virus aggregates. Virus release, as seen by light microscopy, gave the appearance of occurring by a 'budding' process whereby part of the cell would swell and subsequently rupture or break away. Infectivity studies indicated that approx. 80% of newly replicated virus was released extracellularly in aggregates which required disruption to maximize infectious virus yield. Trypsin did not enhance virus infectivity. Of the six EHDV isolates used in this study each isolate was characterized by its own maximum yield obtained after several serial passages in cell culture.

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Section: Electron Microscopic Observationssupporting

confidence: 44%

“…Effect of trypsin on EHD V infectivity. Known amounts of EHDV were treated with 5 /~g/ml and 75/~g/ml trypsin according to the method of Svehag et al (1966). EHDV-infected cell cultures were also treated with 5/~g/ml trypsin according to the method of Almeida et al (1978).…”

Section: Cells and Virusmentioning

confidence: 99%

Replication and Release of Epizootic Haemorrhagic Disease Virus in BHK-21 Cells

Lewanczuk

Yamamoto

1982

Journal of General Virology

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“…Previous reports on the pH stability of bluetongue virus indicate that the virus is extremely labile at acidic pH's with the critical pH being about 6.3 [4,17,25]. Our results were in agreement with these observations.…”

Section: Discussionsupporting

confidence: 93%

“…Bluetongue virus has been reported to be resistant to ether [4,22,25], chloroform [25] and deoxycholate [4,22,25]. Our observations confirmed these findings for Ibaraki virus.…”

Section: Discussionsupporting

confidence: 90%

Ibaraki Virus, an Agent of Epizootic Disease of Cattle Resembling Bluetongue

Inaba

Tanaka

Ishii

et al. 1970

Japanese Journal of Microbiology

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Replication of Ibaraki virus was not inhibited by 5‐iodo‐2′‐deoxyuridine, indicating that the virus is an RNA virus. The virus was resistant to ether, chloroform and deoxycholate, sensitive to trypsin, very labile at acidic pH but stable at pH 6.4 or higher, and was resistant to repeated freezing and thawing. The virus was readily inactivated at 56 C or higher, was fairly stable at 37 C, and very stable at 4 C, while it rapidly lost infectivity when stored frozen at —20 C. The virus was readily sedimented by centrifugation at 40 000Xg for 60 min. It readily passed through membrane filters of 200 mμ pore size, passed through 100 μfilters but only with some titer loss and did not through 50 mμ filters. In these tests, the bluetongue virus used as a control behaved in the same manner as Ibaraki virus. These findings provide additional evidence for the similarity of Ibaraki virus to bluetongue virus which had been previously demonstrated on the basis of seasonal incidence, symptomatology and pathology of the diseases caused by these viruses and the behavior of the viruses in cell cultures, embryonated eggs and laboratory animals. The present study, however, provided no evidence for any serological relation between these two viruses. More Information is needed to reach a final decision on the classification of Ibaraki virus, particularly regarding the morphology of the virion, the doublestrandedness of the viral RNA and other basic features.

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“…However, after elution with 3M KSCN and 100 mM Glycine HCl, pH 3.0, elutes were found to be positive for BTV but the infectivity of the virus was completely lost as no CPE was observed in BHK-21 cell upon inoculation. This may be due to inactivation of BTV at low pH [15] and denaturation of viral proteins by KSCN, a chaotropic agent [12]. Therefore, 4M MgCl 2 with 75 mM HEPES, pH6.5 has been used as an elution buffer for purification of infective BTV by IAC.…”

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confidence: 99%

Purification of infective bluetongue virus particles by immuno-affinity chromatography using anti-core antibody

2016

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An immuno-affinity chromatography technique for purification of infective bluetongue virus (BTV) has been descried using anti-core antibodies. BTV anti-core antibodies (prepared in guinea pig) were mixed with cell culture-grown BTV-1 and then the mixture was added to the cyanogens bromide-activated protein-A Sepharose column. Protein A binds to the antibody which in turn binds to the antigen (i.e. BTV). After thorough washing, antigen-antibody and antibody-protein A couplings were dissociated with 4M MgCl 2 , pH6.5. Antibody molecules were removed by dialysis and virus particles were concentrated by spin column ultrafiltration. Dialyzed and concentrated material was tested positive for BTV antigen by a sandwich ELISA and the infectivity of the chromatography-purified virus was demonstrated in cell culture. This method was applied for selective capture of BTV from a mixture of other viruses. As group-specific antibodies (against BTV core) were used to capture the virus, it is expected that virus of all BTV serotypes could be purified by this method. This method will be helpful for selective capture and enrichment of BTV from concurrently infected blood or tissue samples for efficient isolation in cell culture. Further, this method can be used for small scale purification of BTV avoiding ultracentrifugation.

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Sensitivity of bluetongue virus to lipid solvents, trypsin and pH changes and its serological relationship to arboviruses

Cited by 21 publications

References 20 publications

Replication and Release of Epizootic Haemorrhagic Disease Virus in BHK-21 Cells

Replication and Release of Epizootic Haemorrhagic Disease Virus in BHK-21 Cells

Ibaraki Virus, an Agent of Epizootic Disease of Cattle Resembling Bluetongue

Purification of infective bluetongue virus particles by immuno-affinity chromatography using anti-core antibody

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