Effects of fresh garlic extract on Candida albicans Sessile cells, biofilms and biofilm associated genes, Flo-8 and Ndt80

Dave Thomas, Joshua Lee, S. Choo, H. Rao, P. Chong
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引用次数: 1

Abstract

Candida-associated nosocomial infections are a persistent problem which has been steadily increasing over the years. The emergence of antimicrobial resistant strains has narrowed the spectrum of effective drugs that can be utilised and indicated the need for alternative therapeutics. Garlic is a spice often studied for its unique characteristics. Most of its antimicrobial properties have been attributed to the presence of allicin, which makes up more than 70% of thiosulfinates extracted from garlic. The present study aimed to determine the effects of fresh garlic extract (FGE) towards the expression of two major biofilm genes, Flo-8 and Ndt80 produced by C. albicans that enable the morphogenesis of yeast from planktonic cells to biofilm.  Minimum concentration required for FGE to inhibit C. albicans was determined through agar well and disc diffusion assay. Effect of FGE towards preformed C. albicans biofilms was evaluated at ¼× and ½× inhibitory concentrations of FGE and Amphotericin B, respectively by XTT assay. RNA from FGE-treated C. albicans was extracted, reverse transcribed and analysed by Real-Time Polymerase Chain Reaction (RT-PCR) to determine FGE effects towards the expression of Flo-8 and Ndt80. Inhibitory concentration of FGE was determined to be 100 mg/mL. Both genes appeared to be upregulated in the presence of FGE with a higher upregulation noted with Flo-8 (67.34 and 30.20-folds) when compared against Ndt80 (5.55 and 1.79-folds) at 5 mg/mL and 10 mg/mL, respectively. It is surmised that C. albicans upregulated these genes as a survival mechanism in the presence of FGE. The findings from this study indicate that FGE could inhibit C. albicans growth and biofilms. Further research is required to determine the minimum inhibitory concentration (MIC) through broth microdilution and to understand the mechanism behind this phenomenon before it can be utilised as a potential therapeutic to combat Candida-associated nosocomial infections.
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鲜大蒜提取物对白色念珠菌无柄细胞、生物膜及生物膜相关基因Flo-8和Ndt80的影响
念珠菌相关的医院感染是一个持续存在的问题,多年来一直在稳步增加。抗菌素耐药菌株的出现缩小了可用有效药物的范围,并表明需要替代疗法。大蒜是一种香料,因其独特的特性而经常被研究。它的大部分抗菌特性归因于大蒜素的存在,大蒜中提取的硫代亚硫酸盐中有70%以上是大蒜素。本研究旨在确定新鲜大蒜提取物(FGE)对白色念珠菌产生的两个主要生物膜基因Flo-8和Ndt80表达的影响,这些基因使酵母从浮游细胞向生物膜的形态发生。通过琼脂孔和圆盘扩散试验确定了FGE抑制白色念珠菌所需的最低浓度。在FGE和两性霉素B的抑制浓度分别为¼×和½×时,采用XTT法评价FGE对预制白色念珠菌生物膜的影响。提取经FGE处理的白念珠菌RNA,进行逆转录,RT-PCR检测FGE对fl -8和Ndt80表达的影响。测定FGE的抑制浓度为100 mg/mL。FGE存在时,这两个基因都出现了上调,与Ndt80(5.55和1.79倍)相比,FGE在5 mg/mL和10 mg/mL时的上调幅度更高,其中Flo-8的上调幅度分别为67.34和30.20倍。据推测,白色念珠菌在FGE存在下上调这些基因是一种生存机制。本研究结果表明,FGE能抑制白色念珠菌的生长和生物膜的形成。需要进一步的研究来确定通过肉汤微量稀释的最低抑制浓度(MIC),并了解这一现象背后的机制,然后才能将其用作对抗念珠菌相关医院感染的潜在治疗方法。
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