Electron microscopic studies of bovine viral diarrhea virus in tissues of diseased calves and in cell cultures

1982 ◽  
Vol 71 (1) ◽  
pp. 57-74 ◽  
Author(s):  
H. Bielefeldt Ohmann ◽  
B. Bloch
2017 ◽  
Vol 45 (Supplementary 1) ◽  
pp. 314-322
Author(s):  
Eman M. Hafz ◽  
Ali A.A. Salama ◽  
Adel M. Soliman ◽  
Mohammed EL-Bakry A. Ismail ◽  
Gamelat K. Farag ◽  
...  

Author(s):  
R. Alain ◽  
S. Cardin ◽  
L. Berthiaume ◽  
J. Lecomte

Bovine viral diarrhea virus (BVDV) is an economically important pathogen of cattle throughout the world, causing a wide range of clinical syndromes. Virion is spherical, 40-70 nm in diameter, with an envelope tightly applied to a spherical nucleocapsid 25-35 nm in diameter with icosahedral symmetry (figure 1 and 2) (Matthews, 1979) . It is presently included in the Togavlridae family, genus Pneumovirus but Collett et al. (1988) have proposed to group this genus in the Flaviviridae family. In the present study, two strains of BVDV were compared by electron microcospy ultrathin sections: one cytopathogenic, NADL strain, and the other non-cytopathogenic, NY-1 strain. These were Inoculated on Madney-Darby Bovine Kidney (MDBK) cells to a multiplicity of infection (m.o.i) between 3,0 and 5,0. Control and virus infected cells were fixed with glutaraldehyde (2,5% in cacodylate buffer) at 24 , 48, 72 and 96 h following incubation at 37°C. Cells were postfixed with osmium tetroxide and embedded in Vestopal.The NADL-CP strain infected cells showed vacuolization of the endoplasmic reticulum, ribosomal activity in the cytoplasm and appearance of virus particles 48h after infection (figure 3). At 72h, an expensive vacuolization was apparent (figure 4). Maximum viral production was seen 96 h after infection (figure 5). From 24 to 72 h post infection, the NY-l-NCP strain of BVD infected cells showed normal vacuolization like in control cells. They produced weak vacuolization with few particles, not apparent before 96h after infection (figure 6). Viral particles of 45-50 nm in endoplasmic reticulum vacuoles were in general homogenous in size while those in smooth membranes, probably of cellular origin, heterogenous in size.


2012 ◽  
Vol 153 (1) ◽  
pp. 77-81 ◽  
Author(s):  
L. V. Uryvaev ◽  
A. V. Dedova ◽  
L. V. Dedova ◽  
K. S. Ionova ◽  
N. A. Parasjuk ◽  
...  

1989 ◽  
Vol 1 (1) ◽  
pp. 29-33 ◽  
Author(s):  
Leon N. D. Potgieter ◽  
Kenny V. Brock

A cytopathic strain of bovine viral diarrhea virus (BVDV) was purified from infected cell culture fluids by isopycnic density-gradient centrifugation. Genomic RNA was extracted and tailed with adenine residues at the 3′ end with poly-A polymerase. Double-stranded complementary DNA (cDNA) was synthesized, using the poly-A-tailed RNA as a template and oligo-dT as a primer, and then cloned into the pUC9 plasmid. Virusspecific cDNA sequences, varying in length from 0.5 to 2.5 kilobases (kb), were obtained. One BVDV-specific sequence of cloned cDNA, 1.1 kb in length and with an internal Pst I restriction endonuclease cleavage site, was selected for use as a probe. The cloned cDNA insert was removed from the plasmid either with or without flanking plasmid sequences and labeled with 32P-nucleotides by nick translation for use as hybridization probes for BVDV. The performance of probes of smaller fragments of the insert was compared to that of the intact sequence in hybridization assays. In addition, 2 methods of specimen preparation were compared to establish optimum parameters for hybridization. The hybridization assay was 10–100 times more sensitive than infectivity assays for BVDV in infected cell cultures. Freezing of specimens reduced by lo-fold the sensitivity of hybridization for BVDV target sequences. The probes prepared from the cloned cDNA hybridized with all cytopathic and noncytopathic BVDV strains tested but not with uninfected cell cultures, cellular ribosomal RNA, bovine coronavirus, bluetongue virus, or bovine adenovirus 3. Probes prepared with native plasmid DNA did not hybridize with BVDV or uninfected cell cultures. Probes of 1.1 kb, 0.6 kb, and 0.5 kb hybridized with equal sensitivity to target sequences in the dot blot system. The presence of flanking plasmid sequences did not improve the hybridization performance of the probes. Hybridization results with specimens prepared by RNAsin and heat treatment were similar to those obtained after Nonidet P-40 and formaldehyde treatment.


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