3.3 Degranulation of mast cells
To detect mast cells at the attachment sites of ticks, we stained the tissue sections with toluidine blue. Toluidine blue staining revealed that there was no significant difference in the presence of mast cells in the tick bite site at different feeding periods (day 0 to day 5) of the primary infestations. In the primary infestation, very few degranulated mast cells were found in the surrounding tissues of tick attachment. Also, no significant increase in the infiltration and degranulation of mast cells was noticed in the secondary and tertiary infestations (Supplementary Figure 1).