Scanning Electron Microscopy of African Swine Fever Virus in Tissue Culture

Abstract

Previous studies of African swine fever virus (ASFV) development and its hemadsorption property by transmission electron microscopy showed virus emergence from the cells (1) and the attachment of infected cells to red cells (2). The scanning electron microscope has been used in the present study to examine the surfaces of infected leucocytes and Vero cells. Leucocytes prepared from swine blood were grown in buffer in Leighton tubes containing glass microscope cover slips and infected with ASFV Lisbon '60 strain prepared from tissue culture. After 1 hr. incubation at 37°C, the inoculum fluid was removed and replaced with F15 balanced salt solution maintenance fluid and 10% foetal bovine serum. The Vero cell cultures were grown and infected in a similar manner. Observations were made starting 18 hrs. after infection.The cover slips were washed gently with Sorenson's buffer pH 7.3, fixed in 2% Glutaraldehyde in Sorenson's buffer and post-fixed in 2% osmic acid in Sorenson's buffer. Dehydration with graded alcohols was followed by immersion in amyl acetate.