Inhibition Effect of Garlic (Allium sativum) Extract on Streptococcus sanguinis Biofilm Formation Involving Bacterial Motility Mechanism
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Abstract
Introduction: Streptococcus sanguinis is a primary colonizer in oral biofilm formation, often implicated in infective endocarditis. Methods to control oral biofilm formation are yet to be developed. Garlic (Allium sativum) has shown antimicrobial activities against many pathogen species. We sought to observe the potential of garlic extract to inhibit bacterial adherence to hydroxyapatite (HA) discs as a model of the tooth surface. Methods: Susceptibility of S. sanguinis ATCC 10556 to garlic extract was examined by minimum inhibitory concentration (MIC) test using broth microdilution method. For bacterial adherence assay, saliva-coated HA discs were incubated with various concen- trations of extract then stimulated with S. sanguinis ATCC 10556 suspension. Adherent bacteria were stained with 0.1% crystal violet and measured at 595 nm using a microplate reader. A qualitative method to test bacterial motility was performed using Motility Indole Ornithine (MIO) medium. Results: The result of minimum inhibitory concentration test showed that MIC value for garlic ethanolic extract was at a concentration of 625 μg/ml. Moreover, garlic extract inhibited bacterial adherence to HA discs starting at concentration of 62.5 μg/ml. The inhibition of bacterial motility can be observed, indicated as limited the diffused growth of bacteria closer to the inoculating line. Obser- vation using SEM confirmed these results. Conclusion: This present study suggest that garlic extract has the ability to inhibit S. sanguinis adherence to HA discs involving inhibition of bacterial motility, with the optimal concentration being 500 μg/ml.
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Kreth J, Merritt J, Qi F. Bacterial and Host Interactions of Oral Streptococci. DNA and Cell Biology. 2009; 28(8): 397-403
Okahashi N, Nakata M, Sakurai A, Terao Y, Hosino T, Isoda K, Sumitomo T, et al. Pili of oral Streptococcus sanguinis bind to fibronectin and contribute to cell adhesion. Biochem Biophys Res Commun. 2010; 391:1192-06
Diaz PI, Kolenbrander PE. Subgingival Biofilm Communities in Health and Disease. Rev Clin Periodoncia Implantol Rehabil. Oral. 2009; 2(3):187-92
Pratt LA, Kolter R. Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol. Microbiol. 1998; 30:285–93
Ge X, Kitten T, Chen Z, Lee SP, Munro CL, Xu P. Identification of Streptococcus sanguinis genes required for biofilm formation and examination of their role in endocarditis virulence. Infect Immun. 2008; 76(6):2551-9
Pelicic V. Type IV pili: e pluribus unum? Mol Microbiol. 2008; 68: 827–37
Westling K. Viridans group Streptococci septicaemia and endocarditis. 2005. Karolinka Instituted, Stockholm
Marsh PD, Moter A, Devine DA. Dental plaque biofilms: communities, conflict and control. Periodontology 2000. 2011; (55):16-35
Cutler RR, Wilson P. Antibacterial activity of a new, stable, aqueous extract of allicin against methicillin-resistant Staphylococcus aureus. Br. J. Biomed.Sci. 2004; 61:71–4
Bachrach G, Jamil A, Naor R, Tal G, Ludmer Z, Steinberg D., Garlic Allicin as a Potential Agent for Controlling Oral Pathogens. J. Med. Food. 2014; 14: 1338-43
Hutomo S, Susilowati H, Agustina D, Asmara W. Analysis of Anti-S. sanguinis IgY Ability to Inhibit Streptococcus sanguinis Adherence. Dent J (Maj Ked Gi). 2018;51(1):33-6
Bramanti I, Ngatijan, Purwono S. The acceleration of garlic (Allium sativum L) ethanolic extract on gingival wound healing process in Wistar rats. J. Med. Sci. 2013;45(2): 51-60
Yamaguchi M, Terao Y, Ogawa T, Takahashi T, Hamada S, Kawabata S. Role of Streptococcus sanguinis sortase A in bacterial colonization. Microbes and Infection. 2006; 8: 2791-96
Siswomihardjo W, Sunarintyas S, Tontowi AE. The Effect of Zirconia in Hydroxyapatite on Staphylococcus epidermidis Growth. Int J Biomater, 2012; 432372
Raja AF, Ali F, Khan IA, Shawi AS, Arora DS. Acettyl-11=keto-β-boswellic acid (AKBA); targeting oral cavity pathogens. BMC Res Notes. 2011;4:406
Liu Y, Tortora G, Ryan ME, Lee HM, Golub LM. Potato dextrose agar antifungal susceptibility testing for yeasts and molds: evaluation of phosphate effect on antifungal activity of CMT-3. Antimicrob. Agents. Chemother. 2002;46(5):1455-61
Setiawati S, Nuryastuti T, Ngatijan N, Mustofa M, Jumina J, Fitriastuti D. In Vitro Antifungal Activity of (1)-N-2-Methoxybenzyl-1, 10-phenanthrolinium Bromide against Candida Albicans and Its Effect on Membrane Integrity. Mycobiology. 2017;45(1): 25-30
Wu X, Santos RR, Fink-Gremmels J. Analyzing the antibacterial effects of food ingredients: model experiments with allicin and garlic extracts on biofilm formation and viability of Staphylococcus epidermidis. Food science & Nutrition. 2015;3(2): 158-68
O’May C, Tufenkji N. The Swarming Motility of Pseudomonas aeruginosa Is Blocked by Cranberry Proanthocyanidins and Other Tannin- Containing Materials. Appl. Environ. Microbiol. 2011;77(9):3061-67
Gurung I, Spielman I, Davies MR, Lala R, Gaustad P, Biais N, et al. Functional analysis of an unusual type IV pilus in the Gram-positive Streptococcus sanguinis. Molecular Microbiology. 2015;99:380- 92
Berry JL, Pelicic V. Exceptionally widespread nano-machines composed of type IV pilins: the prokaryotic Swiss Army knives. FEMS Microbiol Rev. 2015;39:134–54
Santas J, Almajano MP, Carbo R. Antimicrobial and antioxidant activity of crude onion (Allium cepa L.) extracts. Int J Food Sci Technol. 2010;45:403-409
Ankri S., and Mirelman D. Antimicrobial properties of allicin from garlic. Microbes Infect. 1999;1:125– 129
Kyu HK. Antimicrobial properties of allium species. Current opinion in Biotechnology. 2012;23:142-47
Fujisawa H, Watanabe K, Suma K, Origuchi K, Matsufuji H, Seki T, Ariga T. Antibacterial potential of garlic-derived allicin and its cancellation by sulfhydryl compounds. Biosci Biotechnol Biochem. 2009;73:1948-55
Ligthart K, Belzer C, de Vos WM, Tytgat HLP. Bridging Bacteria and the Gut: Functional Aspects of Type IV Pili. Trends in Mycrobiology. 2020;28(5):340-8
Colombo AV, Silva CM, Haffajee A. Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesion. J. Med. Microbiol. 2006;55:609-15
Okahashi N, Nakata M, Terao Y, Isoda K, Sakurai A, Sumitomo T, Yamaguchi M, Kimura RK, Oiki E, Kawabata S, Ooshima T. Pili of oral Streptococcus sanguinis bind to salivary amylase and promote the biofilm formation. J. Microbial Pathogenesis. 2011;50:148-54