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Abstract

There are increasing number of reports on Candida albicans developing resistance to available anti-fungal drugs. Thus, there is an urgent need to discover new agents for treatment of candidiasis. The alcoholic extracts of Orthoshipon aristatus have been shown to exhibit antifungal activity against C. albicans by using the agar diffusion and broth microdilution methods. However, the underlying mechanisms of anti-C. albicans effect of O. aristatus have not been well understood. This study was aimed to evaluate the cytotoxic and anti-biofilm effects of the n-hexane and ethanol extracts of purple and white varieties of O. aristatus leaves and branches against C. albicans. The effect of n-hexane and ethanol extract against C. albicans growth was carried out by crystal violet viability assay. IC50 values of the most active extract, and nystatin and fluconazole as positive controls were also determined by the crystal violet assay. Evaluation of the anti-biofilm effect was performed by treating C. albicans with the extracts at adhesion, development, and biofilm maturation stages using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The n-hexane extract of the purple variety of O. aristatus leaves demonstrated the strongest cytotoxic activity against C. albicans amongst the tested extracts, with an IC50 value of 0.67 µg/mL. The extract also showed strong anti-biofilm effect as fluconazole, with pronounced inhibition at the adhesion stage and less activity at the biofilm development and maturation stages. These results suggested that the n-hexane extract of the purple variety of O. aristatus leaves could be explored for discovery and development of anti-C. albicans agent.

References

Atanasov, A.G., Waltenberger, B., Pferschy-Wenzig, E.M., Linder, T., Wawrosh, C., Uhrin, P., et al. (2015). Discovery and resupply of pharmacologically active plant-derived natural products: A review. Biotechnology Advances, 33:1582-1161.

Ashraf, K., Sultan S, Adam A. Orthoshipon stamineus Benth, is an outstanding food medicine: Review of phytochemical and pharmacological activities. Journal of Pharmacy and Bioallied Sciences, 10:109-118.

Freiesleben, S.H,, Jager, A.K. (2014). Correlation between plant secondary metabolites and their anti-fungal mechanisms–A review. Medicinal & Aromatic Plants, 3:154-160.

Hossain, M.A., Ismail, Z., Rahman, A., Kang, S.C. (2008). Chemical composition and anti-fungal properties of the essential oils and crude extracts of Orthosiphon stamineus Benth. Industrial Crops and Products, 27:328–334.

Kadosh D., Lopez-Ribot, J.L. (2013). Candida albicans: Adapting to succeed. Cell Host & Microbe, 14:483-485

Khan, M., Ahmed, J., Gul, A., Ikram, A., Lalani, F.K. (2018). Anti-fungal susceptibility testing of vulvovaginal Candida species among women attending antenatal clinic in tertiary care hospitals of Peshawar. Infection and Drug Resistance, 11:447-456.

Lohse, M., Gulati, M., et al. Development and regulation of single- and multi-species Candida albicans biofilms. Nature Reviews Microbiology,16:19–31.

Lortholary, O., Desnos-Ollivier, M., Sitbon, K., Fontanet, A., Bretagne, S., Dromer, F. (2011). Recent exposure to caspofungin or fluconazole influences the epidemiology of candidemia: a prospective multicenter study involving 2,441 patients. Antimicrobial Agents and Chemotherapy, 55:532–538.

Lyckender, I.M., Malterud, K.E. (1996). Lipophillic flavonoids from Orthosiphon spicatus prevent oxidative inactivation of 15-lipoxygenase. Prostaglandins, Leukotrienes & Essential Fatty Acids, 54:239-246.

Maubon, D., Garnaud, C., Calandra, T., Sanglard, D., Cornet, M. (2014). Resistance of Candida spp. to anti-fungal drugs in the ICU: where are we now?. Intensive Care Medicine, 40 (9): 1241–1255.

McCall, A.D., Pathirana, R.U., Prabhakar, A., Cullen, P.J., Edgerton, M. (2019). Candida albicans biofilm development is governed by cooperative attachment and adhesion maintenance proteins. Biofilms and Microbiomes, 5: 21.

Mohamadi, J., Motaghi, M., Panahi, J., Havasian, M.R., Delpisheh, A., Azizian, M., et al. (2014). Anti-fungal resistance in candida isolated from oral and diaper rash candidiasis in neonates. Bioinformation, 10:667-670.

Mukesh, K.Singh., Bina, Gidwani., Anshita, Gupta., Hemant, Dhongade., Chanchal Deep Kaur., Pranita, P. Kashyap., D.K. Tripathi. (2015). A Review of the medicinal plants of genus orthosiphon (Lamiaceae). International Journal of Biological Chemistry, 9:318-331.

Oxman, D.A., Chow, J.K., Frendl, G., Hadley, S., Hershkovitz, S., Ireland, P., et al. (2010). Candidaemia associated with decreased in vitro fluconazo,le susceptibility: is Candida speciation predictive of the susceptibility pattern?. Journal of Antimicrobial Chemotherapy, 65:1460–1465.

Raekiansyah, M., Buerano, C.C., Luz, M.A.D., Morita, K. (2018). Inhibitory effect of the green tea molecule EGCG against dengue virus infection. Archives of Virology, 163:1649-1655.

Rahmasari, R., Haruyama, T., Charyasriwong, S., Nishida, T., Kobayashi, N. (2017). Antiviral activity of aspalathus linearis against human influenza virus. Natural Product Communications, 12:599-602.

Raut, J.S., Chauhan, N.M., Shinde, R.B., Karuppayil, S.M. (2013). Inhibition of planktonic and biofilm growth of Candida albicans reveals novel antifungal activity of caffeine. Journal of Medicinal Plants Research, 7(13):777-782.

Romulo, A., Zuhud, E.A.M., Rondevaldova, J., Kokoska, L. (2018). Screening of in vitro antimicrobial activity of plants used in traditional Indonesian medicine. Pharmaceutical Biology, 56:287-293

Sandai, D., Tabana, Y.M., El Quweini, A., Ayodeji, I.O. (2016). Resistance of Candida albicans biofilm to drugs and the host immune system. Jundishapur Journal of Microbiology, 9:37385.

Shahzad, M., Sherry, L., Rajendran, R., Edwards, C.A., Combet, E., Ramage, G. (2014). Utilising polyphenols for the clinical management of Candida albicans biofilms. International Journal of Antimicrobial Agents, 44:269-273.

Sorgo, A.G., Heilmann, C.J., Dekker, H.L., Bekker, M., Brul, S., de Koster, C.G., et al. (2011). Effects of fluconazole on the secretome, the wall proteome, and wall integrity of the clinical fungus Candida albicans. Eukaryotic cells, 10:1071–1081.

Tezuka, Y., Stampoulis, P., Banskota, A.H., Awale, S., Tran, K.Q., Saiki, I., et al. (2000). Constituents of the vietnamese medicinal plant Orthosiphon stamineus. Chemical and Pharmaceutical Bulletin, 48:1711–1719.

Traba, C., & Jun F. Liang, J.F. (2011). Susceptibility of Staphylococcus aureus biofilms to reactive discharge gases. Biofouling, 27(7): 763–772.

Watanabe, K., Rahmasari, R., Matsunaga, A., Haruyama, T., Kobayashi, N. (2014). Anti-influenza viral effects of honey in vitro: potent high activity of Manuka honey. Archives of Medical Research, 45:359-365.

Zore, G.B., Thakre, A.D., Jadhav, S., Karuppayil, S.M. (2011). Terpenoids inhibit Candida albicans growth by affecting membrane integrity and arrest of cell cycle. Phytomedicine, 18:1180-1190.

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