Antiviral activity and mode of action of extracts from neem seed kernel against duck plague virus in vitro1.

Four fractions obtained from alcohol extracts of neem (Azadirachta indica) seed kernel by column chromatography were investigated for antivirus activity against the duck plague virus (DPV) in vitro. Duck embryo fibroblasts (DEF) infected with DPV were treated with the neem seed kernel extracts, and the effect of antivirus was judged by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide colorimetric method assay and direct immunofluorescence assay. The mode of action was tested by the plaque reduction assay. The results showed that fractions 1 to 3 were inactive. The median inhibitory concentration (IC(50)) of fraction 4 was 10.9 μg/mL and inhibited the virus protein expression in the direct immunofluorescence assay. In the plaque reduction assay, fraction 4 could significantly reduce the number of plaques compared with the negative control (P < 0.01) in all modes of action. This study indicated that the fourth fraction obtained from neem seed kernel could improve the viability of infected cells, and reduce the cytopathic effects caused by DPV and the amount of the virus protein expressed in virus-infected cells. The antiviral activity works in the whole process of virus infecting the normal cells.