Porcine epidemic diarrhea virus (PEDV) was adapted to serial propagation in Vero cell cultures by adding trypsin to the medium. PEDV-infected cells showed a distinct cytoplasmic fluorescence when examined by a fluorescent-antibody-staining technique. Cytopathic effects, such as vacuolation, formation of syncytia, and fusion of cells, were detected even at passage 1 of the PEDV in Vero cells. Once adapted, the virus induced numerous syncytia containing over 100 nuclei. From virus passage 5 on, all cells forming the monolayer were fused and totally destroyed within 24 h after inoculation. Cell culture-grown PEDV had typical coronavirus morphology when viewed by electron microscopy. Attempts to propagate PEDV in several primary and secondary fetal porcine cell cultures in the presence or absence of trypsin were unsuccessful.
solution, already known for other compounds of this type,"] can also be observed here; through equal coupling with the three '03Rh nuclei, the isochronous benzene protons give rise to a quartet at 6 = 4.06, JRh." = 0.7 Hz in the 'HNMR spectrum (C,D,) at room temperature, whereas the Cp protons appear as singlet at 6 = 5.11.
Physical inclusion of small molecules in larger structural lattices is well known in the crystalline state and is a common feature of the chemistry of zeolites. In the liquid state, a variety of synthetic macrocyclic molecules are available to complex and contain smaller guest species. An alternative strategy for binding is explored: assembly of cavity-forming structures from small subunits. Encapsulation of small guest molecules such as methane can be achieved with a synthetic structure that assembles reversibly through hydrogen bonding.
Eleven European isolates of bovine herpesvirus 1 (BHV-1), together with two reference virus strains were compared by restriction endonuclease digestion, by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and by their reactivity with a panel of monoclonal antibodies (McAb's). Based on the cleavage pattern of viral DNA with the restriction endonuclease Hind III the strains could be assigned to one of two established major virus types. Analysis by SDS-PAGE of viral polypeptides revealed that four protein species either displayed virus type or subtype specific minor variation of migration characteristics. Of 43 McAb's tested all reacted with all type 1 strains, whereas five antibodies failed to recognize some of the type 2 viruses. The existence of type specific variations among virus specified proteins was further evidenced by the recovery of one McAb recognizing type 1 viruses only. The data show that BHV-1 isolates can be assigned to established virus types according to the SDS-PAGE profile of viral proteins or the selective reactivity with type specific McAb's.
The porcine epidemic coronavirus (PEDV), tentatively classified as a coronavirus, was adapted to Vero cells and a plaque test developed for infectivity titration, allowing us to test the biological and biophysical properties of the virus. Growth kinetics showed peak titers of 10(5.5) plaque-forming units ml-1 15 h after infection. Filtration experiments and electron microscopy revealed a particle diameter between 100 and 200 nm. The buoyant density of the virus was 1.18. The particle lost its infectivity on treatment with lipid solvents. Virus replication could not be inhibited by 5-iodo-2'-deoxyuridine. PEDV was moderately stable at 50 degrees C, but heat sensitivity was not altered by divalent cations. At 4 degrees C, the virus was stable between pH 5.0 and 9.0, but at 37 degrees C stability was restricted to the pH range 6.5-7.5. Viral infectivity was not impaired by ultrasonication or by multiple freezing and thawing. PEDV was not neutralized by transmissible gastroenteritis virus antiserum. On the basis of the tests carried out, PEDV is a pleomorphic, enveloped RNA virus with a particle diameter of approximately 150 nm and a buoyant density of 1.18. Infectivity depends on the presence of trypsin, and infected cells show a tendency to fuse and to form syncytia. All of these properties, as well as its physicochemical characteristics, allow PEDV to be classified as a coronavirus.
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