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.

Highlights

  • Acanthamoeba sp. is one of free-living amoeba (FLA), which is an opportunistic pathogen widely spread in the environment [1]

  • Recent studies showed that Acanthamoeba T4 genotypes are the main cause of Acanthamoeba keratitis (AK), a severe progressive corneal inflammation which can lead to blindness [3]

  • Cellulase Concentration and Incubation Time Optimization. Both samples from AK patients were identified as Acanthamoeba castellani T4 genotype, and sample from biosphere was identified as Acanthamoeba lenticulata T5 genotype

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Summary

Introduction

Acanthamoeba sp. is one of free-living amoeba (FLA), which is an opportunistic pathogen widely spread in the environment [1]. According to 18s rRNA sequence, Acanthamoeba sp. Genus can be divided into 20 genotypes, T1-T20 [2]. Recent studies showed that Acanthamoeba T4 genotypes are the main cause of Acanthamoeba keratitis (AK), a severe progressive corneal inflammation which can lead to blindness [3]. Acanthamoeba keratitis is often associated with contact lenses misuse [4]. Complications towards contact lenses use have arisen with the increase of contact lenses user over time; around 4-10% of contact lens users underwent mild irritation to blindness [5]. Each year around 80.000 people suffer from eye disease due to contact lens use [6]. Contact lens users have increased by around 15% each year in Indonesia. The keratitis occurrence was rarely reported, because of undistinguishable clinical manifestation between fungal and bacterial infection [7]

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