Experiment 2: Recovery from sterile sand, beach sand, swamp mud,
and forest soil on macrophages
The collected soils were tested along with two controls: blood-only and
blood mixed with 6 grams of sterile sea sand. All soils and controls
were spiked with 1.2 mL of ASFV-positive blood with a titer of 6.00
log10 HAD50/mL and three replicates per
condition. The forest soils were extremely dry, so 12 ml of media were
added to all samples for virus isolation. We continued experiment 2 and
subsequent experiments at room temperature (25°C), since we had not seen
big differences between soil stored at 4°C or 25°C in experiment 1. We
limited our testing to 14 days in experiment 2 as it seemed unlikely
that live virus would be detected beyond one week. In addition to the
protocol described in experiment 1, we also used a dedicated kit for DNA
extraction from soil (DNeasy PowerSoil; Qiagen, Hilden, Germany) from
0.25 grams of all matrices and time points.