Potential anti-Acanthamoeba and anti-adhesion activities of Annona muricata and Combretum trifoliatum extracts and their synergistic effects in combination with chlorhexidine against Acanthamoeba triangularis trophozoites and cysts


Plants with medicinal properties have been used in the treatment of several infectious diseases, including Acanthamoeba infections. The medicinal properties of Cambodian plant extracts; Annona muricata and Combretum trifoliatum were investigated against Acanthamoeba triangularis. A total of 39 plant extracts were evaluated and, as a result, 22 extracts showed positive anti-Acanthamoeba activity. Of the 22 extracts, 9 and 4 extracts showed antiAcanthamoeba activity against trophozoites and cysts of A. triangularis, respectively. The minimum inhibitory concentration of A. muricata and C. trifoliatum extracts against trophozoites and cysts was 500 and 1,000 mu g/mL, respectively. The combination of A. muricata at 1/4xMIC with chlorhexidine at 1/8xMIC demonstrated a synergistic effect against trophozoites, but partial synergy against cysts. A 40% reduction in trophozoites and 60% of cysts adhered to the plastic surface treated with both extracts at 1/2xMIC were noted comparing to the control (P < 0.05). Furthermore, a reduction of 80% and 90% of trophozoites adhered to the surface was observed after pretreatment with A. muricata and C. trifoliatum extracts, respectively. A 90% of cysts adhered to the surface was decreased with pre-treatment of A. muricata at 1/2xMIC (P < 0.05). A 75% of trophozoites and cysts from Acanthamoeba adhered to the surface were removed after treatment with both extracts at 4xMIC (P < 0.05). In the model of contact lens, 1 log cells/mL of trophozoites and cysts was significantly decreased post-treatment with both extracts compared to the control. Trophozoites showed strong loss of acanthopodia and thorn-like projection pseudopodia, while cysts demonstrated retraction and folded appearance treated with both extracts when observed by SEM, which suggests the potential benefits of the medicinal plants A. muricata and C. trifoliatum as an option treatment against Acanthamoeba infections.




Multidisciplinary Sciences


Mitsuwan, W; Sin, C; Keo, S; Sangkanu, S; Pereira, MD; Jimoh, TO; Salibay, CC; Nawaz, M; Norouzi, R; Siyadatpanah, A; Wiart, C; Wilairatana, P; Mutombo, PN; Nissapatorn, V

nossos autores


This work was supported by the project entitled Medicinal underexploited Thai native plant against Acanthamoeba, Leishmania donovani, and Plasmodium falciparumToward South East Asia collaboration initiative (Grant No. WUBG0202564) supported by The Royal Patronage of Her Royal Highness Princess Maha Chakri Sirindhorn, Walailak University, Thailand.

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