Experiment 4: Recovery of live virus with WSL-adapted CD2v-deleted
ASFV Kenya and disinfection treatments
A virus stock was prepared by mixing 160 mL of supernatant from WSL
cells infected with WSL-adapted CD2v-deleted ASFV Kenya virus with 500
mL of defibrinated whole blood from a domestic swine resulting in a
final ASFV titer of 6.00 log10TCID50/mL.
A 2 mL volume of the spiked blood was used to inoculate 6 g beach sand
and commercial potting soil at room temperature. A blood-only tube and
sterile sand were again included as controls. The protocol was followed
as described above, with the omission of sonication as the samples were
too numerous to sonicate in the 3-hour intervals between the first three
collections. All sample conditions were completed with 3 replicates.
Samples were treated with 3.5% and 7.5% calcium hydroxide or 3.5% and
7.5% citric acid (by weight of soil). After each soil matrix was
inoculated, the powdered disinfectants were added, vortexed and
incubated for 1 or 3 hours at room temperature.