The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n - 3), eicosapentaenoic acid (EPA, 20:5n - 3) and arachidonic acid (ARA, 20,4n -
6) on lipase enzyme activity in pearl spot larvae was studied. Rotifers were enriched with lipid emulsion containing nine different percentage of PUFA (DHA - 5%, 10%, 15%, of total fatty acids, EPA - 5%, 10%, 15%, of total fatty acids, ARA - 5%, 10%, 15% of total fatty acids). And control emulsion containing only olive oil (no DHA, EPA, and ARA). Larvae were fed differently enriched rotifers thrice a day. Initial lipase enzyme analysis was carried out on the first day of rearing and after 15 days of rearing larvae fed with enriched rotifers (Brachionus plicatilis). The maximum lipase activity was recorded in treatments T3 (5.64b±0.01 U/mg protein), T6 (5.76a±0.01 U/mg protein) and T9 (5.54c±0.00 U/mg protein) with inclusion level of 15% - DHA, 15% - EPA, 15% - ARA. Hence the present study could draw a conclusion that the inclusion level of (15%) DHA, (15%) EPA, (15%) ARA will, in turn, increases the enzyme secretion in pearl spot larvae and showed a highly significant difference among the treatments (p<0.05).