Phenoloxidase Activity in Hemolymph of Plasmodium-Melanizing and Nonmelanizing Strains of Anopheles gambiae

ABSTRACT Phenoloxidase (PO) activity was compared in two strains of the mosquito, Anopheles gambiae , one of which melanotically encapsulates and kills malaria parasites ( Plasmodium spp.) and a second which permits normal development of these parasites. The enzyme assay, based on the formation of dopachrome from L-dopa, was linear with respect to enzyme concentration and time and was as inhibitable by phenylthiourea. There were no significant differences in the K m or V max of hemolymph samples taken from 3 days old naive mosquitoes of each strain. An age course demonstrated that PO activity was slightly higher in the refractory strain on Days 1 and 2 after emergence but decreased to similar stable levels in both strains by Day 3. PO activity differed in the two strains following an uninfected blood meal. While PO activity significantly increased by 48 h post blood feeding in Plasmodium -susceptible strain, activity remained level or slightly decreased in the Plasmodium -refractory strain. Finally, protease inhibitors and proteases were tested for effects on PO activity. Leupeptin, TLCK, and TPCK all inhibited PO activity in both strains. PO in both strains was activated by bovine trypsin but not by bovine chymotrypsin.