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      Hibiscus sabdariffa Extract Inhibits Adhesion, Biofilm Initiation and Formation in Candida albicans

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          Abstract

          Microbial biofilms act as reservoirs for pathogenic sessile microbes which reside inside the three dimensional matrix of the biofilm, and are thus protected against anti-microbial drugs. Most of the anti-microbial drugs fail to completely abolish the biofilm associated infections. In the present study, we provide evidence of Hibiscus sabdariffa (Hs) extract having possible anti-microbial activity, with emphasis on Candida albicans biofilm. The Hs extract was shown to be effective against C. albicans pre-formed biofilm at 3.125 mg/ml and was able to inhibit the hyphae initiation and adherence of cells. Furthermore, Hs extract was able to reduce the C. albicans load in C. elegans by effectively killing the Candida cells thereby reducing the viable colony count and effectively increasing the lifespan of worms. The percentage of viable hatched progeny of worms exposed to Hs extract (both at conc. 1.5 mg/ml and 6.25 mg/ml), was also comparable to that of the control untreated eggs. The Hs extract was also found to be significantly effective against fluconazole resistant C. albicans isolated from patients. Thus, we, for the first time, propose Hs extract as a prospective drug candidate and substitute for eradicating pre-formed biofilm and inhibiting the growth of C. albicans .

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          Most cited references20

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          Environmental sensing and signal transduction pathways regulating morphopathogenic determinants of Candida albicans.

          Candida albicans is an opportunistic fungal pathogen that is found in the normal gastrointestinal flora of most healthy humans. However, under certain environmental conditions, it can become a life-threatening pathogen. The shift from commensal organism to pathogen is often correlated with the capacity to undergo morphogenesis. Indeed, under certain conditions, including growth at ambient temperature, the presence of serum or N-acetylglucosamine, neutral pH, and nutrient starvation, C. albicans can undergo reversible transitions from the yeast form to the mycelial form. This morphological plasticity reflects the interplay of various signal transduction pathways, either stimulating or repressing hyphal formation. In this review, we provide an overview of the different sensing and signaling pathways involved in the morphogenesis and pathogenesis of C. albicans. Where appropriate, we compare the analogous pathways/genes in Saccharomyces cerevisiae in an attempt to highlight the evolution of the different components of the two organisms. The downstream components of these pathways, some of which may be interesting antifungal targets, are also discussed.
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            Candida Species Biofilms’ Antifungal Resistance

            Candida infections (candidiasis) are the most prevalent opportunistic fungal infection on humans and, as such, a major public health problem. In recent decades, candidiasis has been associated to Candida species other than Candida albicans. Moreover, biofilms have been considered the most prevalent growth form of Candida cells and a strong causative agent of the intensification of antifungal resistance. As yet, no specific resistance factor has been identified as the sole responsible for the increased recalcitrance to antifungal agents exhibited by biofilms. Instead, biofilm antifungal resistance is a complex multifactorial phenomenon, which still remains to be fully elucidated and understood. The different mechanisms, which may be responsible for the intrinsic resistance of Candida species biofilms, include the high density of cells within the biofilm, the growth and nutrient limitation, the effects of the biofilm matrix, the presence of persister cells, the antifungal resistance gene expression and the increase of sterols on the membrane of biofilm cells. Thus, this review intends to provide information on the recent advances about Candida species biofilm antifungal resistance and its implication on intensification of the candidiasis.
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              In vitro growth and analysis of Candida biofilms.

              Evaluation of fungal biofilm formation can be performed using several techniques. In this protocol, we describe methods used to form Candida biofilms on three different medical device substrates (denture strips, catheter disks and contact lenses) to quantify them and to evaluate their architecture and drug susceptibility. Biofilm formation involves adhesion of fungal cells to pretreated substrates, followed by growth in medium. Formed biofilms are quantified by determining their metabolic activity and dry weight, whereas their gross morphology and architecture are evaluated using fluorescence microscopy, scanning electron microscopy and confocal scanning laser microscopy techniques. Susceptibility of biofilms is determined by comparing their metabolic activity in the presence of antifungal agents with that in their absence. The methods described here can be completed in a typical laboratory with minimum involvement of software. Evaluation of the growth of fungal biofilms and their analyses can be completed using the described methods in approximately 15 d.
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                Author and article information

                Contributors
                (View ORCID Profile)
                Journal
                Indian Journal of Microbiology
                Indian J Microbiol
                Springer Science and Business Media LLC
                0046-8991
                0973-7715
                March 2020
                November 4 2019
                March 2020
                : 60
                : 1
                : 96-106
                Article
                10.1007/s12088-019-00835-9
                7000594
                32089579
                dd955914-54e8-45b0-8f8d-f8a2dcd0d29c
                © 2020

                http://www.springer.com/tdm

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