Malaysian Journal of Analytical Sciences Vol 23 No 5 (2019): 812 - 817

APPLICATION OF DIRECT FLUORESCENCE-BASED LIVE/DEAD STAINING FOR ASSESSMENT OF ANTIFUNGAL ACTIVITY OF COCONUT OIL AGAINST Candida albicans

(Aplikasi Pewarnaan Hidup/Mati Berasaskan Pendarfluor Langsung untuk Penilaian Aktiviti Antikulat Minyak Kelapa terhadap Candida albicans)

Nor Izzah Mukhtar¹, Zurainie Abllah²*, Azrul Naim Mohamad¹, Intan Azura Shahdan³, Kamariah Long⁴, Ummi Aqilah Haron¹

¹Department of Biotechnology, Kulliyyah of Science

²Department of Pediatric Dentistry and Dental Public Health, Kulliyyah of Dentistry

³Department of Biomedical Sciences, Kulliyyah of Allied Health Sciences

International Islamic University Malaysia, 25200 Kuantan, Pahang, Malaysia

⁴Biotechnology Research Centre,

MARDI Headquarters, Serdang, P.O. Box 12301, 50774 Kuala Lumpur, Malaysia

*Corresponding author: drzura@iium.edu.my

Received: 16 January 2019; Accepted: 3 August 2019

Abstract

Candida albicans is one of the common causes for oral candidiasis worldwide. The proliferation of antifungal-resistant C. albicans has become a major concern. This study was carried out to evaluate activated virgin coconut oil (AVCO) and crude extract of virgin coconut oil (VCO) as new antifungal agents for treatment of oral candidiasis. C. albicans viability was determined using LIVE/DEAD bacterial viability kit. C. albicans cells were grown in yeast peptone dextrose (YPD) broth overnight. The fungus was treated with AVCO and VCO at the concentration of minimum fungicidal concentration (MFC) of 6.24 mg/mL and incubated for three different time points (1, 2, and 3 h). To evaluate the viability of C. albicans, SYTO 9 and propidium iodide (PI) staining were used and the cells were observed using fluorescence microscopy. C. albicans treated with AVCO showed more dead cells compared to cells treated with VCO. The data indicate that exposure of C. albicans to AVCO was the most inhibitory to its growth (p < 0.01).

Keywords: LIVE/DEAD staining, `antifungal activity, Candida albicans, virgin coconut oil

Abstrak

Candida albicans ialah salah satu penyebab kepada kandidiasis mulut di seluruh dunia. Malahan, pembiakan C. albicans rintang antikulat semakin mendapat perhatian. Kajian ini dijalankan untuk menilai kesan-kesan minyak kelapa dara teraktif (AVCO) dan ekstrak minyak kelapa dara (VCO) dalam pencarian agen antikulat yang baharu untuk rawatan kandidiasis mulut. Kebolehhidupan sel C. albicans ditentukan dengan penggunaan kit daya kebolehhidupan bakteria HIDUP/MATI. C. albicans dibiakkan semalaman di dalam kultur cecair yis pepton dekstrosa (YDP). Kulat tersebut dirawat dengan minyak kelapa teraktif dan ekstrak minyak kelapa dara pada kepekatan fungisidal minimum (MFC) sebanyak 6.24 mg/mL dan dieramkan pada tiga masa yang berbeza (1, 2, dan 3 jam). Untuk menilai kebolehhidupan C. albicans, pewarnaan SYTO 9 dan propidium iodida digunakan dan sel dilihat dengan menggunakan mikroskop pendarfluor. C. albicans yang telah dirawat dengan minyak kelapa dara teraktif menunjukkan lebih banyak sel yang mati berbanding sel yang dirawat dengan ekstrak minyak kelapa dara. Data menunjukkan minyak kelapa dara teraktif adalah paling berkesan untuk menghalang pembiakan C. albicans (p < 0.01).

Kata kunci: pewarnaan HIDUP/MATI, aktiviti rintang kulat, C. albicans, minyak kelapa dara

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