Pouzolzia indica Methanolic Extract Fraction 2 and Povidone-Iodine Induced Changes in the Cyst of Acanthamoeba spp.: Light and Electron Microscopic Studies
Jantima Roongruangchai DDS, PhD*, Tichaporn Sookkua MS*, Tanawan Kummalue MD, PhD**, Kosol Roongruangchai MS, MD***
Affiliation : * Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand ** Department of Clinical Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand *** Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
Objective : To compare the minimal cysticidal concentration (MCC) between Pouzolzia indica methanolic
extract fraction 2 and Povidone-Iodine (PVP-I) on the Acanthamoeba cyst and to illustrate the morphological
changes of the cyst after being treated by light and electron microscopies.
Material and Method: Acanthamoeba spp were isolated from patients with Acanthamoeba keratitis and
cultured on a non-nutrient agar plate (NNA) seeded with heat killed Escherichia coli (NNA-E.coli) at 37°C
for 7 days, adjusted to a final concentration of 104cysts/ml. Several concentrations of PVP-I and fraction 2 of
Pouzolzia indica methanolic extract were tested to find the minimal cysticidal concentrations (MCC) of both
agents, at these concentrations there was no viable cyst which was confirmed by no excystment after further
incubation for 7 days. The cysts were prepared for routine transmission and scanning electron microscopic
studies.
Results : Structural damages of the treated cysts by MCC of PVP-I and fraction 2 of Pouzolzia indica methanolic
extract showed a series of damages. Starting from shrinkage, destruction or rupture of the cyst walls and
opercula, withdrawal of the cytoplasm or edema cyst by the outside solution passed through the damaged wall
caused a decrease in wrinkle ridges of the ectocyst. Then the cyst was ripped and torn into small pieces
Concluison: MCC of PVP-I solution and the fraction 2 of Pouzolzia indica methanolic extract were 0.04% and
1:4, respectively. The structural damages were somewhat similar, such as the shrinkage, ruptured cyst wall
and opercula, edema and end by breaking up of the cyst wall and degeneration of the inside cytosol. Pouzolzia
indica may be modified as an effective disinfectant solution for a contact lens case if the active ingredients are
more purified.
Keywords : Pouzolzia indica Benn., Acanthamoeba spp., Povidone-iodine
