American Journal of Infectious Diseases and Microbiology
ISSN (Print): 2328-4056 ISSN (Online): 2328-4064 Website: http://www.sciepub.com/journal/ajidm Editor-in-chief: Maysaa El Sayed Zaki
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American Journal of Infectious Diseases and Microbiology. 2014, 2(5), 122-130
DOI: 10.12691/ajidm-2-5-5
Open AccessArticle

N-acetylcysteine Inhibits and Eradicates Candida albicans Biofilms

El-Baky Rehab Mahmoud Abd1, , Dalia Mohamed Mohamed Abo El Ela1 and Gamal Fadl Mamoud Gad1

1Department of Microbiology, Faculty of Pharmacy, Minia University, Minia, Egypt

Pub. Date: November 04, 2014

Cite this paper:
El-Baky Rehab Mahmoud Abd, Dalia Mohamed Mohamed Abo El Ela and Gamal Fadl Mamoud Gad. N-acetylcysteine Inhibits and Eradicates Candida albicans Biofilms. American Journal of Infectious Diseases and Microbiology. 2014; 2(5):122-130. doi: 10.12691/ajidm-2-5-5

Abstract

N-acetylcysteine (NAC) is used in the treatment of chronic bronchitis that attributed to its mucus dissolving properties. Its ability to reduce biofilm formed by different types of bacteria was proven previously in many studies. Therefore we examined its effect on C. albicans biofilms by testing its effect alone and in combination with ketoconazole using Tissue culture plate assay method (TCP). NAC effects on C. albicans morphology and the texture of biofilms were determined using Scanning electron microscope (SEM). It was found that the inhibitory effect of NAC was concentration dependent. NAC reduced C. albicans adherence by ≥32.8% while ketoconazole reduced adherence by ≥25% in comparison to control. Also, it showed higher disruptive effect (50-95%) than ketoconazole (22-80.7%) on mature biofilms. Using NAC and ketoconazole in combination, a significant inhibitory effect (P<0.01) on both adherence and mature biofilms (54-100%) was seen. NAC reduced the amount of biofilm mass in all tested Candida in concentrations at which their growth was not affected. NAC and ketoconazole combinations showed complete eradication to mature biofilms formed in most of the tested strains. NAC can inhibit C. albicans growth, inhibit dimorphism, which is an important step in biofilm formation, and change the texture of the formed biofilms, what makes NAC an interesting agent to be used as an inhibitor for biofilm formation by C. albicans.

Keywords:
antifungal mature biofilm mucolytics adherence SEM

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References:

[1]  Banerjee SN, Emori TG, Culver DH, Gaynes RP, Jarvis WR, Horan T, Edwards JR, Tolson J, Henderson T, Martone WJ. Secular trends in nosocomial primary bloodstream infections in the United States, 1980-1989. National Nosocomial Infections Surveillance System. Am J Med 1991, 91: 86S-8.
 
[2]  Williams DW, Kuriyama T, Silva S, Malic S, Lewis MAO. Candida biofilms and oral candidosis: treatment and prevention. Periodontology 2000 2011, 55: 250-265.
 
[3]  Marsh PD, Martin M. “Oral fungal infections,” in Oral Microbiology, Churchill Livingstone, Edinburgh, UK, 2009:166-179.
 
[4]  Nobile, CJ, Schneider HA, Nett JE, Sheppard DC, Filler SG, Andes DR, Mitchell AP. Complementary adhesin function in C. albicans biofilm formation. Curr Biol 2008, 18: 1017-1024.
 
[5]  Baillie GS, Douglas LJ. Candida biofilms and their susceptibility to antifungal agents. Methods Enzymol, 1999, 310: 644-656.
 
[6]  Nobile CJ, Nett JE, Andes DR, Mitchell AP. Function of Candida albicans adhesion Hwp1 in biofilm formation. Eukaryotic Cell 2006, 5:1604-1610.
 
[7]  Nett JE, Lepak AJ, Marchillo K, Andes DR. Time course global gene expression analysis of an in vivo Candida biofilm. J Infect Dis 2009, 200: 307-313.
 
[8]  Donlan RM, Costerton JW. Biofilms: Survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 2002, 15: 167-93.
 
[9]  Fux CA, Costerton JW, Stewart PS, Stoodley P. Survival strategies of infectious biofilms. Trends Microbiol 2005, 13: 34-40.
 
[10]  Nikawa H, Nishimura H, Makihira S, et al. Effect of serum concentration on Candida biofilm formation on acrylic surfaces. Mycoses 2000, 43: 139-143.
 
[11]  Hawser SP, Douglas LJ. Resistance of Candida albicans biofilms to antifungal agents in vitro. Antimicrob Agents Chemother 1995, 399: 2128-31.
 
[12]  Mermel LA., Farr BM, Sherertz RJ, et al.. Infectious Diseases Society of America American College of Critical Care Medicine Society for Healthcare Epidemiology of America. Guidelines for the management of intravascular catheter-related infections. Clin Infect Dis 2001, 329: 1249-72.
 
[13]  Pappas PG, Kauffman CA, Andes D, et al. Infectious Diseases Society of America. Clinical practice guidelines for the management of candidiasis. Clin Infect Dis 2009, 485: 503-35.
 
[14]  Riise GC, Qvarfordt I, Larsson S, Eliasson V and Andersson BA. Inhibitory effect of N-acetylcysteine on adherence of Streptococcus pneumoniae and Haemophilus influenzae to human oropharyngeal epithelial cells in vitro. Respiration 2000, 67: 552-558.
 
[15]  Stey C, Steurer J, Bachmann S, Medici TC, and Tramer MR. The effect of oral N-acetylcysteine in chronic bronchitis: a quantitative systematic review. Eur. Respir. J. 2000, 16: 253-262.
 
[16]  Blanco MT, Blanco J, Sanchez-Benito R, Perez-Giraldo C, Moran F J, Hurtado C, and Gomez-Garcia A C. Incubation temperatures affect adherence to plastic of Candida albicans by changing the cellular surface hydrophobicity. Microbios 1997, 89: 23-28.
 
[17]  Sheffner AL. The reduction in vitro in viscosity of mucoprotein solutions by a new mucolytic agent, N-acetyl-L-cysteine. Ann. N.Y. Acad. Sci. 1963, 106: 298-10.
 
[18]  Perez-Giraldo C, Rodriguez-Benito A, Moran FJ, Hurtado C, Blanco MT, Gómez-García AC. Influence of N-acetylcysteine on the formation of biofilm by Staphylococcus epidermidis. Journal of Antimicrobial Chemotherapy. 1997, 39:643-6.
 
[19]  Olofsson AC, Hermansson M, Elwing H. N-acetyl-L-cysteine affects growth, extracellular polysaccharide production, and bacterial biofilm formation on solid surfaces. Appl Environ Microbiol., 2003, 69: 4814-22.
 
[20]  Schmitt-Andrieu L and Hule C. Alginates of Pseudomonas aeruginosa: a complex regulation of the pathway of biosynthesis. C.R. Acad. Sci. Ser. III 1996, 19:153-160.
 
[21]  Benson HC. Microbiological Application: Laboratory Manual in General Microbiology, 11th ed., McGram-Hill Higher Education, Sanfrancisco, 2002, pp.168.
 
[22]  Christensen GD, Simpson WA, Younger JA, Baddour LM, Barrett FF, Melton, DM, et al. Adherence of coagulase negative Staphylococci to plastic tissue cultures: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 1985, 22: 996-1006.
 
[23]  Pfaller, M. A., C. Grant, V. Morthland, and J. Rhine-Chalberg. 1994. Comparative evaluation of alternative methods for broth dilution susceptibility testing of fluconazole against Candida albicans. J. Clin. Microbiol. 32:506–509.
 
[24]  .Esimone C O, Adiukwu M U, Okonta J M, "Preliminary Antimicrobial Screening of the Ethanolic Extract from the Lichen Usnea subfloridans L,". IJPRD. 1998 3:99-102.
 
[25]  Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA. Biofilm formation by the fungal pathogen Candida albicans. development, architecture, and drug resistance. J Bacteriol 2001, 83: 5385-5394.
 
[26]  Merritt K, Hitchins VM, Brown SA. Safety and cleaning of medical materials and devices. J Biomed Mater Res 2000, 53: 131-136.
 
[27]  Ramage G, VandeWalle K, Wickes BL, Lopez-Ribot JL. Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. Antimicrob Agents Chemother 2001, 45: 2475-2479.
 
[28]  Xiaogang L, Zhun Y and Jianping X. Quantitative variation of biofilms among strains in natural populations of Candida albicans. Microbiology 2003, 149: 353-362.
 
[29]  Soboh F, Khoury AE, Zamboni AC, Davidson D, Mittelman MW. Effects of ciprofloxacin and protamine sulfate combinations against catheter-associated Pseudomonas aeruginosa biofilms. Antimicrob. Agents Chemother 1995, 39: 1281-1286.
 
[30]  Sivasubramanian G, and Sobel JD. Refractory urinary tract and vulvovaginal infection caused by Candida krusei. Int. Urogynecol. J. Pelvic Floor Dysfunct., 2009, 20:1379-138.
 
[31]  Jin Y, Yip HK, Samaranayake YH, Yau JY and Samaranayake LP. Biofilm-Forming Ability of Candida albicans Is Unlikely To Contribute to High Levels of Oral Yeast Carriage in Cases of Human Immunodeficiency Virus Infection J. Clin. Microb 2003, 4170: 2961-2976.
 
[32]  Kuhn DM, George T, Chandra J, Mukherjee PK, and Ghannoum MA. Comparison of biofilms formed by Candida albicans and Candida parapsilosis on bioprosthetic surfaces. Infect. Immun 2002, 70:878-888.
 
[33]  Dominic RM, Shenoy S, Baliga S. Candida biofilms in medical devices: evolving. trends Kathmandu Univ Med J 2007, 5:431-43.
 
[34]  Hoyle BD, Jass J, Costerton JW. The biofilm glycocalyx as a resistance factor. J Antimicrob Chemother 1990, 26: 1-5.
 
[35]  Al-Fattani MA, Douglas LJ. Penetration of Candida biofilms by antifungal agents. Antimicrob Agents Chemother 2004, 48: 3291-7.
 
[36]  Nett JE, Crawford K, Marchillo K, Andes DR. Role of Fks1p and matrix glucan in Candida albicans biofilm resistance to an echinocandin, pyrimidine, and polyene. Antimicrob Agents Chemother 2010, 54: 3505-8.
 
[37]  Gilbert P, Collier PJ, Brown MRW. Influence of growth rate on susceptibility to antimicrobial agents: biofilms, cell cycle, dormancy, and stringent response. Antimicrob. Agents Chemother. 1990, 34:1865-1868.
 
[38]  Harriott MM, Lilly EA, Rodriguez T E, Fidel P L Jr, and Noverr M. C. Candida albicans forms biofilms on the vaginal mucosa. Microbiology, 2010, 156, 3635-3644.
 
[39]  Baillie GS, Douglas LJ. Effect of growth rate on resistance of Candida albicans biofilms to antifungal agents. Antimicrob. Agents Chemother. 1998, 42:1900-1905.
 
[40]  Chebotar IV, Parshikov VV. Investigation of the effect of antimycotics on the Candida biofilms. Scientific and practical journal of obstetrics and gynecology, 2013, 5: 98-100.
 
[41]  El-Feky M A, El-Rehewy M S, Hassan M A, Aboulella H A and Abd El-Baky R M, Gad G F.. Effect of ciprofloxacin and N-acetylcysteine on bacterial adherence and biofilm formation on ureteral stent surfaces. Pol. J Microbiol, Vol. 2009, 58, No 3, 261.267.
 
[42]  Aslam S and Darouiche R. Role of Antibiofilm-Antimicrobial Agents in Control of Device-Related Infections. Int J Artif Organs. 2011, 34: 752-758.
 
[43]  Venkatesh M, Rong L, Raad I, Versalovic J. Novel synergistic antibiofilm combinations for salvage of infected catheters. J Med Microbiol 2009, 58: 936-44.
 
[44]  Ramage G, VandeWalle K, Lopez-Ribot J L and Wickes B L 2002,. The filamentation pathway controlled by the Efg1 regulator protein is required for normal biofilm formation and development in Candida albicans. FEMS Microbiol. Lett. 214:95-100.
 
[45]  Lewis R E, Lo H J, Raad I I and Kontoyiannis D P. Lack of catheter infection by the efg1/efg1 cph1/cph1 double-null mutant, a Candida albicans strain that is defective in filamentous growth. Antimicrob. Agents Chemother. 2002, 46:1153-1155.