Sains Malaysiana 41(5)(2012): 569–572

 

In Vitro Sensitivity Testing of Acanthamoeba Clinical Isolates from Patients with Keratitis against Polyhexamethylene biguanide(PHMB) and Chlorhexidine

(Kajian Sensitiviti Isolat KlinikalAcanthamoeba daripada Pesakit Keratitis SecaraIn Vitro Terhadap Polyhexamethylene biguanide (PHMB) dan Chlorhexidine)

 

 

Noradilah Samseh binti Abdullah

Fakulti Perubatan dan Sains Kesihatan, Universiti Sains Islam Malaysia, Tingkat 13 Menara B Pesiaran MPAJ, Jalan Pandan Utama, 55100 Pandan Indah, Kuala Lumpur, Malaysia

 

Mohamed Kamel Abd. Ghani*

Biomedical Science Programme, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, 50300 Jalan Raja Muda Abdul Aziz, Kuala Lumpur, Malaysia

 

Anisah Nordin, Yusof Suboh & Noraina Ab Rahim

Department of Parasitology & Entomology, Faculty of Medicine, Universiti Kebangsaan Malaysia

50300 Jalan Raja Muda Abdul Aziz, Kuala Lumpur, Malaysia

 

Norazah Ahmad

Division of Bacteriology, Institute for Medical Research , 50588 Jalan Pahang, Kuala Lumpur Malaysia

 

Received: 21 July 2011 / Accepted: 20 October 2011

 

ABSTRACT

Acanthamoeba keratitis is a serious infection of the eye which can result in permanent visual impairment. Therefore this study was performed to evaluate the effectiveness of antimicrobial agents on three Acanthamoeba clinical isolates (HS 6, HKL 95 and HTH 73). Antimicrobial agents used in this study were polyhexamethylene biguanide (PHMB) and chlorhexidine and both were serially diluted. Cyst suspensions from all three strains were tested against the antimicrobial agents, respectively. After 48 h of incubation at 37°C, the suspension was filtered and the filter membrane was placed onto non-nutrient agar plate lawned with heat-killed Escherichia coli. The plates were examined daily under the inverted microscope until day 14 but were negative for Acanthamoeba trophozoites. The presence of trophozoites indicated ineffectiveness of the antimicrobial agents. Both antimicrobial agents used were found to be effective against Acanthamoeba cysts for all the strains tested. PHMB gave minimum cysticidal concentration (MCC) mean value of 4.232 μg/mL and chlorhexidine showed MCC mean value of 3.906 μg/mL. So, from this study, it can be concluded that PHMB and chlorhexidine were effective in killing the tested Acanthamoeba cysts.

 

Keywords: Acanthamoeba; in vitro; sensitivity; polyhexamethylene biguanide; chlorhexidine

 

ABSTRAK

Keratitis Acanthamoeba merupakan infeksi mata yang serius yang boleh menyebabkan kerosakan penglihatan yang teruk. Oleh yang demikian, kajian ini dilakukan untuk mengkaji keberkesanan agen antimikrob terhadap tiga isolat klinikal Acanthamoeba (HS 6, HKL 95 and HTH 73). Agen antimikrob yang digunakan di dalam kajian ini adalah poliheksametilina biguanide (PHMB) dan kloroheksidin yang menjalani pencairan bersiri. Suspensi sista daripada ketiga-tiga strain diuji dengan kedua-dua agen antimikrob. Selepas inkubasi selama 48 jam pada suhu 37°C, campuran suspensi sista dan agen antimikrob tersebut dituras dan membran turasan diletakkan ke atas plat agar bukan nutrien yang dibanjiriEscherichia coli. Kesemua agar tersebut diperiksa setiap hari menggunakan mikroskop terbalik sehingga hari ke 14 tetapi negatif untuk trofozoitAcanthamoeba. Kehadiran trofozoit menunjukkan ketidakberkesanan agen antimikrob. Kedua-dua agen antimikrob yang digunakan didapati berkesan terhadap sistaAcanthamoeba bagi kesemua strain yang diuji. PHMB memberikan nilaiMCC pada kepekatan 4.232 μg/mL manakala chlorhexidine pula menunjukkan nilaiMCC pada kepekatan 3.906 μg/mL. Maka, daripada kajian ini, dapat disimpulkan bahawaPHMB dan kloroheksidina berkesan membunuh sistaAcanthamoeba yang diuji.

 

Kata kunci: Acanthamoeba; in vitro; kloroheksidine; poliheksametilena biguanide; sensitivitc

REFERENCES

Borazjani, R.N., May, L.L., Noble, A.J., Avery, V.S. & Ahearn, G.D. 2000. Flow cytometry for determination of the efficacy of contact lens disinfecting solutions against Acanthamoeba spp. Appl. Environ. Microbiol. 66:1057-1061.

Elder, M.J., Kilvington, S. & Dart, J.K.G. 1994. A clinicopathologic study of in vitro sensitivity testing and Acanthamoeba keratitis. Investig. Ophthalmol. Vis. Sci. 35:1059-1064.

Goodall, K., Brahma, A. & Ridgway, A. 1996. Acanthamoeba keratitis: masquerading as adenoviral keratitis. Eye 10:643-644.

Kamel A.G.M, Haniza, H. Anisah, N. Yusof, S. Faridah, H. Norhayati, M. & Norazah, A. 2005. More Acanthamoeba keratitis cases in Malaysia. International Medical Journal 12(1): 7-9.

Kilvington, S., Reanne, H., James, B. & John, D. 2002. Activities of therapeutic agents and myristamidopropyl dimethylamine against Acanthamoeba isolates. Antimicrob Agents and Chemother. 46(6): 2007-2009.

Larkin, D.F.P., Kilvington, S. & Dart, J.K.G. 1992. Treatment of Acanthamoeba keratitis with polyhexamethylene biguanide. Ophthalmol 99:185-191.

Ledee, R.D., Iovieno, A., Miller, D., Mandal, N., Diaz, M., Fell, J., Fini, E.M. & Alfonso, C.E. 2009. Molecular Identification of T4 and T5 Genotypes in Isolates from Acanthamoeba Keratitis Patients. J. Clinical. Microbiol. 47(5):1458-1462.

Lee, J., Boo, O., Hee, C. & Hak, Y. 1998. Cysticidal effect on Acanthamoeba and toxicity on human keratocytes by polyhexamethylene biguanide and chlorhexidine. Cornea 26 (6): 736-741

Narasimhan, Sandhya, M.S., Madhavan, Hajib, N. & Lily, T. 2001. Development and application of an in vitro susceptibility test for Acanthamoeba species isolated from keratitis to polyhexamethylene biguanide and chlorhexidine. The J. Cornea and Ext. Dis. 21(2): 203-205

Patel, A. & Hammersmith, K. 2008. Contact lens-related microbial keratitis: recent outbreaks. Curr. Opin. Ophthalmol. 19: 302-306.

Perez-Santoja, J.J., Kilvington, S., Hughes, R., Tufail, A., Matherson, M. & Dart, J.K.G. 2003. Persistently culture positive Acanthamoeba keratitis: In vivo resistance and in vitro sensitivity. Ophtalmol. 110(8): 1593-1600.

Simmons, R. B., Rose, J.L., Crow, A. S. & Ahearn, G.D. 1999. The occurrence and persistence of mixed biofilms in automobile air conditioning systems. Curr. Microbiol. 39: 141-145.

Watson S. 2002. Ocular therapeutic case studies : differential diagnosis and management of microbial keratitis. Opto. (4): 26-31.

 

*Corresponding author; email: mkamal@medic.ukm.my

 

 

 

previous