In vitro susceptibility test for acanthamoeba species isolated from clinical specimens against chlorhexidine, propamidine isethionate, gentamicin and chloramphenicol

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

Research output: Contribution to journalArticle

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Abstract

Introduction: Acanthamoeba keratitis is one of the most severe and potentially sight-threatening ocular parasitic infectious diseases and is recognized as the most challenging among ocular infections. In vitro susceptibility testing of Acanthamoeba isolates may prove beneficial for application of early treatment regimens. This study was conducted to determine the effectiveness of the drugs in therapeutic dose and the minimum cysticidal concentrations (MCCs) of the drugs. 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 3 Acanthamoeba isolates which were isolated from keratitis cases. 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 replication and growth 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 exhibited their cysticidal activities in therapeutic dose but not for gentamicin and chloramphenicol. The minimum cysticidal concentration (MCC) of chlorhexidine ranged from 25 ·g/ml to 50 ·g/ml, propamidine isethionate ranged from 500 ·g/ml to 1000 ·g/ml and gentamicin ranged from 10000 ·g/ml to 20000 ·g/ml. The mean MCC of chlorhexidine, propamidine isethionate and gentamicin on Acanthamoeba isolates was 33.33 ·g/ml, 666.66 ·g/ml and 13333.33·g/ml respectively. Conclusion: The in vitro sensitivity test enables the determination of MCC of drugs on Acanthamoeba isolates and can be used for the screening of new anti-acanthamoebal therapeutic agents.

Original languageEnglish
Pages (from-to)147-149
Number of pages3
JournalInternational Medical Journal
Volume18
Issue number2
Publication statusPublished - Jun 2011

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

Keywords

  • Acanthamoeba
  • In-vitro susceptibility test
  • Malaysia

ASJC Scopus subject areas

  • Medicine(all)

Cite this

In vitro susceptibility test for acanthamoeba species isolated from clinical specimens against chlorhexidine, propamidine isethionate, gentamicin and chloramphenicol. / Abdul Ghani, Mohamed Kamel; Hoon, Shirley Tang Gee; Nordin, Anisah; Hashim, Putri Noradyani Megat; Suboh, Yusof; Rahim, Noraina Abdul.

In: International Medical Journal, Vol. 18, No. 2, 06.2011, p. 147-149.

Research output: Contribution to journalArticle

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abstract = "Introduction: Acanthamoeba keratitis is one of the most severe and potentially sight-threatening ocular parasitic infectious diseases and is recognized as the most challenging among ocular infections. In vitro susceptibility testing of Acanthamoeba isolates may prove beneficial for application of early treatment regimens. This study was conducted to determine the effectiveness of the drugs in therapeutic dose and the minimum cysticidal concentrations (MCCs) of the drugs. 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 3 Acanthamoeba isolates which were isolated from keratitis cases. 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 replication and growth 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 exhibited their cysticidal activities in therapeutic dose but not for gentamicin and chloramphenicol. The minimum cysticidal concentration (MCC) of chlorhexidine ranged from 25 ·g/ml to 50 ·g/ml, propamidine isethionate ranged from 500 ·g/ml to 1000 ·g/ml and gentamicin ranged from 10000 ·g/ml to 20000 ·g/ml. The mean MCC of chlorhexidine, propamidine isethionate and gentamicin on Acanthamoeba isolates was 33.33 ·g/ml, 666.66 ·g/ml and 13333.33·g/ml respectively. Conclusion: The in vitro sensitivity test enables the determination of MCC of drugs on Acanthamoeba isolates and can be used for the screening of new anti-acanthamoebal therapeutic agents.",
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T1 - In vitro susceptibility test for acanthamoeba species isolated from clinical specimens against chlorhexidine, propamidine isethionate, gentamicin and chloramphenicol

AU - Abdul Ghani, Mohamed Kamel

AU - Hoon, Shirley Tang Gee

AU - Nordin, Anisah

AU - Hashim, Putri Noradyani Megat

AU - Suboh, Yusof

AU - Rahim, Noraina Abdul

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N2 - Introduction: Acanthamoeba keratitis is one of the most severe and potentially sight-threatening ocular parasitic infectious diseases and is recognized as the most challenging among ocular infections. In vitro susceptibility testing of Acanthamoeba isolates may prove beneficial for application of early treatment regimens. This study was conducted to determine the effectiveness of the drugs in therapeutic dose and the minimum cysticidal concentrations (MCCs) of the drugs. 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 3 Acanthamoeba isolates which were isolated from keratitis cases. 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 replication and growth 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 exhibited their cysticidal activities in therapeutic dose but not for gentamicin and chloramphenicol. The minimum cysticidal concentration (MCC) of chlorhexidine ranged from 25 ·g/ml to 50 ·g/ml, propamidine isethionate ranged from 500 ·g/ml to 1000 ·g/ml and gentamicin ranged from 10000 ·g/ml to 20000 ·g/ml. The mean MCC of chlorhexidine, propamidine isethionate and gentamicin on Acanthamoeba isolates was 33.33 ·g/ml, 666.66 ·g/ml and 13333.33·g/ml respectively. Conclusion: The in vitro sensitivity test enables the determination of MCC of drugs on Acanthamoeba isolates and can be used for the screening of new anti-acanthamoebal therapeutic agents.

AB - Introduction: Acanthamoeba keratitis is one of the most severe and potentially sight-threatening ocular parasitic infectious diseases and is recognized as the most challenging among ocular infections. In vitro susceptibility testing of Acanthamoeba isolates may prove beneficial for application of early treatment regimens. This study was conducted to determine the effectiveness of the drugs in therapeutic dose and the minimum cysticidal concentrations (MCCs) of the drugs. 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 3 Acanthamoeba isolates which were isolated from keratitis cases. 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 replication and growth 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 exhibited their cysticidal activities in therapeutic dose but not for gentamicin and chloramphenicol. The minimum cysticidal concentration (MCC) of chlorhexidine ranged from 25 ·g/ml to 50 ·g/ml, propamidine isethionate ranged from 500 ·g/ml to 1000 ·g/ml and gentamicin ranged from 10000 ·g/ml to 20000 ·g/ml. The mean MCC of chlorhexidine, propamidine isethionate and gentamicin on Acanthamoeba isolates was 33.33 ·g/ml, 666.66 ·g/ml and 13333.33·g/ml respectively. Conclusion: The in vitro sensitivity test enables the determination of MCC of drugs on Acanthamoeba isolates and can be used for the screening of new anti-acanthamoebal therapeutic agents.

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