Effect of Cellulase Enzyme Treatment on Cyst Wall Degradation of Acanthamoeba sp.

Abstract

Aim. The goal of this study is to know the potential of cellulase in the degradation of cyst wall Acanthamoeba sp. Methods. Sample of Acanthamoeba sp. obtained from isolate collection of Department of Parasitology FKUI of which two samples come from patient and one sample is from environment. All three samples were cultured using non-nutrient agar (NNA) media and identified by PCR and sequencing. The concentration of cellulase concentration used was 50 U, 100 U, 150 U, 200 U, 250 U, and 300 U with the incubation time used being 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours. Furthermore, treatment results with the most optimum concentration and incubation time were observed by using SEM to see changes in the surface of the walls of the cyst. A cysticidal test was performed to determine the effectiveness cysticidal action of disinfectant solution, cellulase, and the combination of disinfectant solution and cellulase in killing Acanthamoeba sp. cyst assessed by their viability value. Results. The most optimal cellulase concentration in killing Acanthamoeba sp. cysts was 300 U with an incubation time of 24 hours. Percentage of viability of Acanthamoeba sp. which was exposed to a disinfectant solution for 24 hours was 95%, cellulase alone for 24 hours 75%, and the combination of cellulase and disinfectant solution for 24 hours 25%. Conclusions. Cellulase is capable of degrading Acanthamoeba sp. cyst wall. Optimal cellulase concentration in degrading Acanthamoeba sp. cyst wall is 300 U with an optimal incubation time being 24 hours. The addition of cellulase to the disinfectant solution has the potential to increase the effectiveness of the disinfectant solution because cellulase is capable of degrading the cyst wall allowing the disinfectant solution to enter and kill Acanthamoeba sp. cysts.

Bioactive Compounds of Ziziphus spina-christi Seeds Extract and Cellulase Enzyme Attenuates the Growth of Acanthamoeba polyphaga Isolated From Contact Lenses

“…Furthermore, treatment with extract and cellulase combined caused morphological changes in cyst walls such as shrinkage and irregularly shaped cysts. This is consistent with Lazuana et al (2019), who observed that a mixture of cellulase and disinfectant solution led to shrunken and irregular shaped cysts. In addition, a combination of CHX and cellulase can disrupt cyst wall structure and enhance the efficacy of marketed contact lens disinfectants against Acanthamoeba castellanii trophozoites and cysts in vitro (Abjani et al, 2017).…”

The Microscopic Detection and Identification of Acanthamoeba Spp. In Water Samples Following Enrichment by in Vitro-Cultivation Under Two Distinct Temperature Conditions.

“…Based on morphological observations using an inverted microscope, 20 samples were identified as positive for Acanthamoeba spp. and PCR confirmed that 18 of these 20 samples were true positives, results that contrast with some previous studies (Khezri et al 2016;Lazuana et al 2019). The difference between morphological identification and PCR is probably due to amoebae other than Acanthamoeba spp.…”

Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes

“…This morphological change may be a sign of cell death, or it may be a protective mechanism of the parasite under stress conditions. To support this finding, an earlier study by Tisha Lazuana and her colleagues revealed that a perfectly round shape and a decreased size of cyst were observed after exposure to cellulose enzyme and disinfectant solution, and this led to Acanthamoeba cell death [ 53 ]. However, the characterization of cell death mechanism by P. harmala extract is of interest to further verify this association before any conclusion could be made.…”