Light and electron microscopic studies of the structural damages of acanthamoeba cysts by pouzolzia indica methanolic extract fraction 4 solution.

Abstract
Objective : This study was to investigate the morphological changes and ultrastructural damages of the Acanthamoeba cysts after treated with the minimal cysticydal concentration (MCC) of Pouzolzia indica methanolic extract fraction 4 solution by light and electron microscopies. Materials and Methods : Acanthamoeba cyst which prepared followed the method of Roongruangchai K15,16 were adjusted to the final concentration of 104 cysts/ml and treated with 1:4 soltion of fraction 4 Pouzolzia indica methanolic extract, the Thai medicinal plant. which was the minimal cysticidal concentration (MCC) 16. The cysts then, were centrifuged and the pellets were prepared for light, scanning and transmission electron microscopies. Result : Light micrographs showed the cytoplasmic clumping and some empty cyst wall. Transmission electron microscopy showed the ruptures of both ectocyst and the endocyst wall with some opercula damages. The cytoplasm aggregated and clumped. Scanning electron microscopy show steps of damage which started from shrinkage and collapsed of the cyst walls, then the cysts began to bulge and swell resulted in decreasing in the wrinkles of the cyst walls. The cytoplasmic contents drain out from the cyst wall and finally the cyst walls were ripped and torn into small pieces. Conclusion : Pouzolzia indica methanolic extract fraction 4 at the MCC of 1:4 caused structural damages to the Acanthamoeba cyst including shrinking of the cyst walls with the ruptured of the ectocyst, endocyst and operculum, edema and break out into pieces. The active ingredients of this Thai medicinal plant should be further studied as this can eventually be one of the regimen for the treatment of the Acanthamoeba keratitis or as a solution for cleaning the contact lens or contact lens case solution.
Description
Keywords
Pouzolzia indica, Acanthamoeba spp., TEM, SEM, LM
Citation
Roongruangchai Jantima, Sookkua Tichaporn, Kummalue Tanawan, Roongruangchai Kosol. Light and electron microscopic studies of the structural damages of acanthamoeba cysts by pouzolzia indica methanolic extract fraction 4 solution. Siriraj Medical Journal, 2012 Jan; 64(suppl): 68-71.