Susceptibility of acanthamoeba species isolated from environmental specimens to chlorhexidine, propamidine isethionate, gentamicin and chloramphenicol

An in-vitro study in Malaysia

Shirley Tang Gee Hoon, Mohamed Kamel Abdul Ghani, Anisah Nordin, Putri Noradyani Megat Hashim, Yusof Suboh, Noraina Abdul Rahim, Norazah Ahmad

Research output: Contribution to journalArticle

Abstract

Introduction: Acanthamoeba keratitis is known as one of the most painful and progressive sight-threatening ocular parasitic infectious diseases. If not diagnose early and treated aggressively, it may cause severe ocular inflammation and visual loss. In vitro studies on the susceptibility of Acanthamoeba isolates may prove beneficial for application of early treatment regimens. The aims of the present study were to determine the effectiveness of the drugs in therapeutic dose and the minimum cysticidal concentrations (MCCs) of the drugs on Acanthamoeba isolates from Malaysia. Materials and Methods: Serial doubling dilutions of chlorhexidine digluconate from 200 ·g/ml to 0.0977 ·g/ml, propamidine isethionate (Brolene ®) from 1000 ·g/ml to 0.4883 ·g/ml and gentamicin from 40000 ·g/ml to 19.5313 ·g/ml were performed in microtiter plate and tested against three Acanthamoeba isolates which were isolated from peat soil. After the exposure of the cysts to the drugs for 24 hours, the cysts were washed free of drugs by centrifugation. The deposit (cysts) was cultured onto nonnutrient agar plates overlaid with heat-killed Escherichia coli. The growth and replication of the trophozoites from cysts exposed to each of the dilutions were observed and recorded microscopically for 14 days to determine the MCC of each drug. The effectiveness of the drugs in therapeutic dose against the cysts was tested directly without any doubling dilutions. Results: Chlorhexidine digluconate and propamidine isethionate (Brolene ®) successfully demonstrated their cysticidal activities in therapeutic dose but not for gentamicin and chloramphenicol. The minimum cysticidal concentration (MCC) of chlorhexidine ranged from 6.25 ·g/ml to 25 ·g/ml, propamidine isethionate was 1000 ·g/ml and gentamicin 10000 ·g/ml. The mean MCC of chlorhexidine, propamidine isethionate and gentamicin on Acanthamoeba isolates was 12.50 ··10.82 ·g/ml, 1000.00 · g/ml and 10000.00 ·g/ml, respectively. From this study, it was clear that there were major differences between the four tested drugs in the in vitro susceptibility test on the Acanthamoeba spp. Conclusion: The present study has indicated that the in vitro susceptibility test has enabled the determination of MCC of drugs on Acanthamoeba isolates. The tested drugs with the MCC values can be considered alone or in combination as potential anti-acanthamoebal therapeutic agents for the treatment of the Acanthamoeba keratitis.

Original languageEnglish
Pages (from-to)325-328
Number of pages4
JournalInternational Medical Journal
Volume18
Issue number4
Publication statusPublished - Dec 2011

Fingerprint

Acanthamoeba
Chlorhexidine
Malaysia
Chloramphenicol
Gentamicins
Cysts
Pharmaceutical Preparations
Acanthamoeba Keratitis
Soil
propamidine isethionate
In Vitro Techniques
Trophozoites
Parasitic Diseases
Therapeutics
Centrifugation
Agar
Communicable Diseases
Hot Temperature
Escherichia coli
Inflammation

Keywords

  • Acanthamoeba
  • In-vitro susceptibility test
  • Malaysia

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Susceptibility of acanthamoeba species isolated from environmental specimens to chlorhexidine, propamidine isethionate, gentamicin and chloramphenicol : An in-vitro study in Malaysia. / Hoon, Shirley Tang Gee; Abdul Ghani, Mohamed Kamel; Nordin, Anisah; Hashim, Putri Noradyani Megat; Suboh, Yusof; Rahim, Noraina Abdul; Ahmad, Norazah.

In: International Medical Journal, Vol. 18, No. 4, 12.2011, p. 325-328.

Research output: Contribution to journalArticle

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abstract = "Introduction: Acanthamoeba keratitis is known as one of the most painful and progressive sight-threatening ocular parasitic infectious diseases. If not diagnose early and treated aggressively, it may cause severe ocular inflammation and visual loss. In vitro studies on the susceptibility of Acanthamoeba isolates may prove beneficial for application of early treatment regimens. The aims of the present study were to determine the effectiveness of the drugs in therapeutic dose and the minimum cysticidal concentrations (MCCs) of the drugs on Acanthamoeba isolates from Malaysia. Materials and Methods: Serial doubling dilutions of chlorhexidine digluconate from 200 ·g/ml to 0.0977 ·g/ml, propamidine isethionate (Brolene {\circledR}) from 1000 ·g/ml to 0.4883 ·g/ml and gentamicin from 40000 ·g/ml to 19.5313 ·g/ml were performed in microtiter plate and tested against three Acanthamoeba isolates which were isolated from peat soil. After the exposure of the cysts to the drugs for 24 hours, the cysts were washed free of drugs by centrifugation. The deposit (cysts) was cultured onto nonnutrient agar plates overlaid with heat-killed Escherichia coli. The growth and replication of the trophozoites from cysts exposed to each of the dilutions were observed and recorded microscopically for 14 days to determine the MCC of each drug. The effectiveness of the drugs in therapeutic dose against the cysts was tested directly without any doubling dilutions. Results: Chlorhexidine digluconate and propamidine isethionate (Brolene {\circledR}) successfully demonstrated their cysticidal activities in therapeutic dose but not for gentamicin and chloramphenicol. The minimum cysticidal concentration (MCC) of chlorhexidine ranged from 6.25 ·g/ml to 25 ·g/ml, propamidine isethionate was 1000 ·g/ml and gentamicin 10000 ·g/ml. The mean MCC of chlorhexidine, propamidine isethionate and gentamicin on Acanthamoeba isolates was 12.50 ··10.82 ·g/ml, 1000.00 · g/ml and 10000.00 ·g/ml, respectively. From this study, it was clear that there were major differences between the four tested drugs in the in vitro susceptibility test on the Acanthamoeba spp. Conclusion: The present study has indicated that the in vitro susceptibility test has enabled the determination of MCC of drugs on Acanthamoeba isolates. The tested drugs with the MCC values can be considered alone or in combination as potential anti-acanthamoebal therapeutic agents for the treatment of the Acanthamoeba keratitis.",
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