Using phospholipases A 2 -specific spectrophotometric assays, it was shown that A. castellanii lysates and their conditioned medium exhibit phospholipase activities. The extracellular levels of PLA 2 detected were significantly reduced compared with the cell-associated enzyme (P<0.05). Sphinganine, a PLA 2 inhibitor showed robust amoebistatic properties but had no effect on the viability of A. castellanii. The potency of sphinganine was demonstrated effectively towards purified PLA 2 derived from porcine pancreas. Using sphinganine, it was observed that PLA 2 is involved in neither binding nor cytotoxicity of the human brain microvascular endothelial cells due to A. castellanii. Unlike as was the case for Dictyostelium amoebae, PLA 2 appeared to be involved in A. castellanii phagocytosis of the fluorescently-labelled polystyrene beads. Horseradish peroxidase was used as a tracer molecule to develop assays to study pinocytosis in A. castellanii. The findings revealed that sphinganine impedes phagocytosis but augments pinocytosis in A. castellanii suggesting distinct nature of processes. A complete understanding of the role of phospholipases in the biology and pathogenesis of A. castellanii infections will determine their potential as therapeutic targets.