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Research article

Food Microbiology and Biotechnology (FMB)  |  Bioactive compounds and Physiological properties

Activity of Allyl Isothiocyanate and Its Synergy with Fluconazole against Candida albicans Biofilms

Jayant Shankar Raut 1, Bhagyashree Shridhar Bansode 2, Ashwini Khanderao Jadhav 2 and Sankunny Mohan Karuppayil 2*

1University Institute of Pharmaceutical Sciences (UIPS), UGC-Centre of Advanced Study, Panjab University, Chandigarh-160014, India, 2School of Life Sciences (DST-FIST & UGC-SAP sponsored), SRTM University, Nanded, 431606 (MH), India

Received: July 29, 2016; Accepted: January 31, 2017

J. Microbiol. Biotechnol. 2017; 27(4): 685-693

Published April 28, 2017 https://doi.org/10.4014/jmb.1607.07072

Copyright © The Korean Society for Microbiology and Biotechnology.

Abstract

Candidiasis involving the biofilms of Candida albicans is a threat to immunocompromised patients. Candida biofilms are intrinsically resistant to the antifungal drugs and hence novel treatment strategies are desired. The study intended to evaluate the anti-Candida activity of allyl isothiocyanate (AITC) alone and with fluconazole (FLC), particularly against the biofilms. Results revealed the concentration-dependent activity of AITC against the planktonic growth and virulence factors of C. albicans. Significant (p <0.05) inhibition of the biofilms was evident at ≤1 mg/ml concentrations of AITC. Notably, a combination of 0.004 mg/ml of FLC and 0.125 mg/ml of AITC prevented the biofilm formation. Similarly, the preformed biofilms were significantly (p <0.05) inhibited by the AITC-FLC combination. The fractional inhibitory concentration indices ranging from 0.132 to 0.312 indicated the synergistic activity of AITC and FLC against the biofilm formation and the preformed biofilms. No hemolytic activity at the biofilm inhibitory concentrations of AITC and the AITC-FLC combination suggested the absence of cytotoxic effects. The recognizable synergy between AITC and FLC offers a potential therapeutic strategy against biofilm-associated Candida infections.

Keywords

Anti-biofilm, Candida, combination, drug resistance, phytochemical, virulence

References

  1. Ganguly S, Mitchell AP. 2011. Mucosal biofilms of Candida albicans. Curr. Opin. Microbiol. 14: 380-385.
    Pubmed PMC CrossRef
  2. Ramage G, Martinez JP, Lopez-Ribot JL. 2006. Candida biofilms on implanted biomaterials: a clinically significant problem. FEMS Yeast Res. 6: 979-986.
    Pubmed CrossRef
  3. Desai JV, Mitchell AP, Andes DR. 2014. Fungal biofilms, drug resistance, and recurrent infection. Cold Spring Harb. Perspect. Med. 1: 10.
    CrossRef
  4. Kojic EM, Darouiche RO. 2004. Candida infections of medical devices. Clin. Microbiol. Rev. 17: 255-267.
    Pubmed PMC CrossRef
  5. Crump JA, Collignon PJ. 2000. Intravascular catheterassociated infections. Eur. J. Clin. Microbiol. Infect. Dis. 19: 1-8.
    Pubmed CrossRef
  6. Mukherjee PK, Chandra J. 2004. Candida biofilm resistance. Drug Res. Updates 7: 301-309.
    Pubmed CrossRef
  7. Fanning S, Mitchell AP. 2012. Fungal biofilms. PLoS Pathog. 8: e1002585.
    Pubmed PMC CrossRef
  8. Srinivasan A, Lopez-Ribot JL, Ramasubramanian AK. 2014. Overcoming antifungal resistance. Drug Disc. Today Technol. 11: 65-71.
    Pubmed PMC CrossRef
  9. Ramage G, Robertson SN, Williams C. 2014. Strength in numbers: antifungal strategies against fungal biofilms. Int. J. Antimicrob. Agents 43: 114-120.
    Pubmed CrossRef
  10. Cowan MM. 1999. Plant products as antimicrobial agents. Clin. Microbiol. Rev. 12: 564-582.
    Pubmed PMC
  11. Cragg GM, Newman DJ. 2013. Natural products: a continuing source of novel drug leads. Biochim. Biophys. Acta 1830:3670-3695.
    Pubmed PMC CrossRef
  12. Lee HS, Kim Y. 2016. Antifungal activity of Salvia miltiorrhiza against Candida albicans is associated with the alteration of membrane permeability and (1,3)-β-D-glucan synthase activity. J. Microbiol. Biotechnol. 26: 610-617.
    Pubmed CrossRef
  13. Raut JS, Karuppayil SM. 2016. Phytochemicals as inhibitors of Candida biofilm. Curr. Pharm. Des. 22: 4111-4134.
    Pubmed CrossRef
  14. Wagner H, Ulrich-Merzenich G. 2009. Synergy research:approaching a new generation of phytopharmaceuticals. Phytomedicine 16: 97-110.
    Pubmed CrossRef
  15. Betts JW, Wareham DW, Haswell SJ, Kelly SM. 2013. Antifungal synergy of theaflavin and epicatechin combinations against Candida albicans. J. Microbiol. Biotechnol. 23: 1322-1326.
    Pubmed CrossRef
  16. Doke SK, Raut JS, Dhawale SC, Karuppayil SM. 2014. Sensitization of Candida albicans biofilms to fluconazole by terpenoids of plant origin. J. Gen. Appl. Microbiol. 60: 163-168.
    Pubmed CrossRef
  17. Wilson AE, Bergaentzlé M, Bindler F, Marchioni E, Lintz A, Ennahar S. 2013. In vitro efficacies of various isothiocyanates from cruciferous vegetables as antimicrobial agents against foodborne pathogens and spoilage bacteria. Food Control 30: 318-324.
    CrossRef
  18. Wu X, Zhou QH, Xu K. 2009. Are isothiocyanates potential anti-cancer drugs? Acta Pharmacol. Sin. 30: 501-512.
    Pubmed PMC CrossRef
  19. Davaatseren M, Hwang JT, Park JH, Kim MS, Wang S, Sung MJ. 2014. Allyl isothiocyanate ameliorates angiogenesis and inflammation in dextran sulfate sodium-induced acute colitis. PLoS One 9: e102975.
    Pubmed PMC CrossRef
  20. Rhee MS, Lee SY, Dougherty RH, Kang DH. 2003. Antimicrobial effects of mustard flour and acetic acid against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium. Appl. Environ. Microbiol. 69:2959-2963.
    Pubmed PMC CrossRef
  21. Shin IS, Masuda H, Naohide K. 2004. Bactericidal activity of wasabi (Wasabia japonica) against Helicobacter pylori. Int. J. Food Microbiol. 94: 255-261.
    CrossRef
  22. Depree JA, Howard TM, Savage GP. 1998. Flavour and pharmaceutical properties of the volatile sulphur compounds of wasabi (Wasabia japonica). Food Res. Int. 31: 329-337.
    CrossRef
  23. Harvey SG, Hannahan HN, Sams CE. 2002. Indian mustard and allyl isothiocyanate inhibit Sclerotium rolfsii. J. Am. Soc. Hort. Sci. 127: 27-31.
  24. Isshiki K, Tokuoka K, Mori R, Chiba S. 1992. Preliminary examination of allyl isothiocyanate vapor for food preservation. Biosci. Biotechnol. Biochem. 56: 1476-1477.
    CrossRef
  25. Nowicki D, Rodzik O, Herman-Antosiewicz A, SzalewskaPałasz A. 2016. Isothiocyanates as effective agents against enterohemorrhagic Escherichia coli: insight to the mode of action. Sci. Rep. 6: 22263.
    Pubmed PMC CrossRef
  26. Abreu AC, Borges A, Mergulhão F, Simões M. 2014. Use of phenyl isothiocyanate for biofilm prevention and control. Int. Biodeterior. Biodegrad. 86: 34-41.
    CrossRef
  27. Borges A, Simões LC, Saavedra MJ, Simões M. 2014. The action of selected isothiocyanates on bacterial biofilm prevention and control. Int. Biodeterior. Biodegrad. 86: 25-33.
    CrossRef
  28. Clinical and Laboratory Standards Institute (CLSI). 2002. Reference method for broth dilution antifungal susceptibility testing of yeast. Approved Standard, M27-A2, 2nd Ed. Clinical and Laboratory Standards Institute, Wayne, PA. USA.
  29. Raut JS, Shinde RB, Chauhan NM, Karuppayil SM. 2014. Phenylpropanoids of plant origin as inhibitors of biofilm formation by Candida albicans. J. Microbiol. Biotechnol. 24:1216-1225.
    Pubmed CrossRef
  30. Raut JS, Shinde RB, Chauhan NM, Karuppayil SM. 2013. Terpenoids of plant origin inhibit morphogenesis, adhesion, and biofilm formation by Candida albicans. Biofouling 29: 87-96.
    Pubmed CrossRef
  31. Liu W, Li LP, Zhang JD, Li Q, Shen H, Chene SM, et al. Synergistic antifungal effect of glabridin and fluconazole. PLoS One 9: e103442.
    Pubmed PMC CrossRef
  32. Shinde RB, Raut JS, Chauhan NM, Karuppayil SM. 2013. Chloroquine sensitizes biofilms of Candida albicans to antifungal azoles. Braz. J. Infect. Dis. 17: 395-400.
    Pubmed CrossRef
  33. Ahmad A, Khan A, Manzoor N, Khan LA. 2010. Evolution of ergosterol biosynthesis inhibitors as fungicidal against Candida. Microb. Pathog. 48: 35-41.
    Pubmed CrossRef
  34. Park HW, Choi KD, Shin IS. 2013. Antimicrobial activity of isothiocyanates (ITCs) extracted from horseradish (Armoracia rusticana) root against oral microorganisms. Biocontrol. Sci. 18: 163-168.
    Pubmed CrossRef
  35. Guiamet PS, Gómez de Saravia SG. 2005. Laboratory studies of biocorrosion control using traditional and environmentally friendly biocides: an overview. Lat. Am. Appl. Res. 35: 295-300.
  36. Ramage G, Rajendran R, Sherry L, Williams C. 2012. Fungal biofilm resistance. Int. J. Microbiol. 2012: 528521.
    Pubmed PMC CrossRef
  37. Verstrepen KJ, Klis FM. 2006. Flocculation, adhesion and biofilm formation in yeasts. Mol. Microbiol. 60: 5-15.
    Pubmed CrossRef
  38. Vila T, Romo JA, Pierce CG, McHardy SF, Saville SP, LopezRibot JL. 2016. Targeting Candida albicans filamentation for antifungal drug development. Virulence 2016: DOI: 10.1080/21505594.2016.1197444.
    CrossRef
  39. Nickerson KW, Atkin AL, Hornby JM. 2006. Quorum sensing in dimorphic fungi: farnesol and beyond. Appl. Environ. Microbiol. 72: 3805-3813.
    Pubmed PMC CrossRef
  40. Cui J, Ren B, Tong Y, Dai H, Zhang L. 2015. Synergistic combinations of antifungals and anti-virulence agents to fight against Candida albicans. Virulence 6: 362-371.
    Pubmed PMC CrossRef
  41. Turgis M, Han J, Caillet S, Lacroix M. 2009. Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi. Food Control 20: 1073-1079.
    CrossRef
  42. Raut JS, Karuppayil SM. 2014. A status review on the medicinal properties of essential oils. Ind. Crops Prod. 62:250-264.
    CrossRef
  43. Xiao D, Srivastava SK, Lew KL, Zeng Y, Hershberger P, Johnson CS, et al. 2003. Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G2/M arrest and inducing apoptosis. Carcinogenesis 24: 891-897.
    Pubmed CrossRef
  44. National Toxicology Program (NTP). 1982. Carcinogenesis bioassay of allyl isothiocyanate (CAS No. 57-06-7) in F344/N Rats and B6C3F1 mice (Gavage study). Natl. Toxicol. Prog. Tech. Rep. Ser. 234: 1-142.
    Pubmed
  45. Zhang Y. 2010. Allyl isothiocyanate as a cancer chemopreventive phytochemical. Mol. Nutr. Food Res. 54: 127-135.
    Pubmed PMC CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2017; 27(4): 685-693

Published online April 28, 2017 https://doi.org/10.4014/jmb.1607.07072

Copyright © The Korean Society for Microbiology and Biotechnology.

Activity of Allyl Isothiocyanate and Its Synergy with Fluconazole against Candida albicans Biofilms

Jayant Shankar Raut 1, Bhagyashree Shridhar Bansode 2, Ashwini Khanderao Jadhav 2 and Sankunny Mohan Karuppayil 2*

1University Institute of Pharmaceutical Sciences (UIPS), UGC-Centre of Advanced Study, Panjab University, Chandigarh-160014, India, 2School of Life Sciences (DST-FIST & UGC-SAP sponsored), SRTM University, Nanded, 431606 (MH), India

Received: July 29, 2016; Accepted: January 31, 2017

Abstract

Candidiasis involving the biofilms of Candida albicans is a threat to immunocompromised
patients. Candida biofilms are intrinsically resistant to the antifungal drugs and hence novel
treatment strategies are desired. The study intended to evaluate the anti-Candida activity of
allyl isothiocyanate (AITC) alone and with fluconazole (FLC), particularly against the
biofilms. Results revealed the concentration-dependent activity of AITC against the planktonic
growth and virulence factors of C. albicans. Significant (p <0.05) inhibition of the biofilms was
evident at ≤1 mg/ml concentrations of AITC. Notably, a combination of 0.004 mg/ml of FLC
and 0.125 mg/ml of AITC prevented the biofilm formation. Similarly, the preformed biofilms
were significantly (p <0.05) inhibited by the AITC-FLC combination. The fractional inhibitory
concentration indices ranging from 0.132 to 0.312 indicated the synergistic activity of AITC
and FLC against the biofilm formation and the preformed biofilms. No hemolytic activity at
the biofilm inhibitory concentrations of AITC and the AITC-FLC combination suggested the
absence of cytotoxic effects. The recognizable synergy between AITC and FLC offers a
potential therapeutic strategy against biofilm-associated Candida infections.

Keywords: Anti-biofilm, Candida, combination, drug resistance, phytochemical, virulence

References

  1. Ganguly S, Mitchell AP. 2011. Mucosal biofilms of Candida albicans. Curr. Opin. Microbiol. 14: 380-385.
    Pubmed KoreaMed CrossRef
  2. Ramage G, Martinez JP, Lopez-Ribot JL. 2006. Candida biofilms on implanted biomaterials: a clinically significant problem. FEMS Yeast Res. 6: 979-986.
    Pubmed CrossRef
  3. Desai JV, Mitchell AP, Andes DR. 2014. Fungal biofilms, drug resistance, and recurrent infection. Cold Spring Harb. Perspect. Med. 1: 10.
    CrossRef
  4. Kojic EM, Darouiche RO. 2004. Candida infections of medical devices. Clin. Microbiol. Rev. 17: 255-267.
    Pubmed KoreaMed CrossRef
  5. Crump JA, Collignon PJ. 2000. Intravascular catheterassociated infections. Eur. J. Clin. Microbiol. Infect. Dis. 19: 1-8.
    Pubmed CrossRef
  6. Mukherjee PK, Chandra J. 2004. Candida biofilm resistance. Drug Res. Updates 7: 301-309.
    Pubmed CrossRef
  7. Fanning S, Mitchell AP. 2012. Fungal biofilms. PLoS Pathog. 8: e1002585.
    Pubmed KoreaMed CrossRef
  8. Srinivasan A, Lopez-Ribot JL, Ramasubramanian AK. 2014. Overcoming antifungal resistance. Drug Disc. Today Technol. 11: 65-71.
    Pubmed KoreaMed CrossRef
  9. Ramage G, Robertson SN, Williams C. 2014. Strength in numbers: antifungal strategies against fungal biofilms. Int. J. Antimicrob. Agents 43: 114-120.
    Pubmed CrossRef
  10. Cowan MM. 1999. Plant products as antimicrobial agents. Clin. Microbiol. Rev. 12: 564-582.
    Pubmed KoreaMed
  11. Cragg GM, Newman DJ. 2013. Natural products: a continuing source of novel drug leads. Biochim. Biophys. Acta 1830:3670-3695.
    Pubmed KoreaMed CrossRef
  12. Lee HS, Kim Y. 2016. Antifungal activity of Salvia miltiorrhiza against Candida albicans is associated with the alteration of membrane permeability and (1,3)-β-D-glucan synthase activity. J. Microbiol. Biotechnol. 26: 610-617.
    Pubmed CrossRef
  13. Raut JS, Karuppayil SM. 2016. Phytochemicals as inhibitors of Candida biofilm. Curr. Pharm. Des. 22: 4111-4134.
    Pubmed CrossRef
  14. Wagner H, Ulrich-Merzenich G. 2009. Synergy research:approaching a new generation of phytopharmaceuticals. Phytomedicine 16: 97-110.
    Pubmed CrossRef
  15. Betts JW, Wareham DW, Haswell SJ, Kelly SM. 2013. Antifungal synergy of theaflavin and epicatechin combinations against Candida albicans. J. Microbiol. Biotechnol. 23: 1322-1326.
    Pubmed CrossRef
  16. Doke SK, Raut JS, Dhawale SC, Karuppayil SM. 2014. Sensitization of Candida albicans biofilms to fluconazole by terpenoids of plant origin. J. Gen. Appl. Microbiol. 60: 163-168.
    Pubmed CrossRef
  17. Wilson AE, Bergaentzlé M, Bindler F, Marchioni E, Lintz A, Ennahar S. 2013. In vitro efficacies of various isothiocyanates from cruciferous vegetables as antimicrobial agents against foodborne pathogens and spoilage bacteria. Food Control 30: 318-324.
    CrossRef
  18. Wu X, Zhou QH, Xu K. 2009. Are isothiocyanates potential anti-cancer drugs? Acta Pharmacol. Sin. 30: 501-512.
    Pubmed KoreaMed CrossRef
  19. Davaatseren M, Hwang JT, Park JH, Kim MS, Wang S, Sung MJ. 2014. Allyl isothiocyanate ameliorates angiogenesis and inflammation in dextran sulfate sodium-induced acute colitis. PLoS One 9: e102975.
    Pubmed KoreaMed CrossRef
  20. Rhee MS, Lee SY, Dougherty RH, Kang DH. 2003. Antimicrobial effects of mustard flour and acetic acid against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium. Appl. Environ. Microbiol. 69:2959-2963.
    Pubmed KoreaMed CrossRef
  21. Shin IS, Masuda H, Naohide K. 2004. Bactericidal activity of wasabi (Wasabia japonica) against Helicobacter pylori. Int. J. Food Microbiol. 94: 255-261.
    CrossRef
  22. Depree JA, Howard TM, Savage GP. 1998. Flavour and pharmaceutical properties of the volatile sulphur compounds of wasabi (Wasabia japonica). Food Res. Int. 31: 329-337.
    CrossRef
  23. Harvey SG, Hannahan HN, Sams CE. 2002. Indian mustard and allyl isothiocyanate inhibit Sclerotium rolfsii. J. Am. Soc. Hort. Sci. 127: 27-31.
  24. Isshiki K, Tokuoka K, Mori R, Chiba S. 1992. Preliminary examination of allyl isothiocyanate vapor for food preservation. Biosci. Biotechnol. Biochem. 56: 1476-1477.
    CrossRef
  25. Nowicki D, Rodzik O, Herman-Antosiewicz A, SzalewskaPałasz A. 2016. Isothiocyanates as effective agents against enterohemorrhagic Escherichia coli: insight to the mode of action. Sci. Rep. 6: 22263.
    Pubmed KoreaMed CrossRef
  26. Abreu AC, Borges A, Mergulhão F, Simões M. 2014. Use of phenyl isothiocyanate for biofilm prevention and control. Int. Biodeterior. Biodegrad. 86: 34-41.
    CrossRef
  27. Borges A, Simões LC, Saavedra MJ, Simões M. 2014. The action of selected isothiocyanates on bacterial biofilm prevention and control. Int. Biodeterior. Biodegrad. 86: 25-33.
    CrossRef
  28. Clinical and Laboratory Standards Institute (CLSI). 2002. Reference method for broth dilution antifungal susceptibility testing of yeast. Approved Standard, M27-A2, 2nd Ed. Clinical and Laboratory Standards Institute, Wayne, PA. USA.
  29. Raut JS, Shinde RB, Chauhan NM, Karuppayil SM. 2014. Phenylpropanoids of plant origin as inhibitors of biofilm formation by Candida albicans. J. Microbiol. Biotechnol. 24:1216-1225.
    Pubmed CrossRef
  30. Raut JS, Shinde RB, Chauhan NM, Karuppayil SM. 2013. Terpenoids of plant origin inhibit morphogenesis, adhesion, and biofilm formation by Candida albicans. Biofouling 29: 87-96.
    Pubmed CrossRef
  31. Liu W, Li LP, Zhang JD, Li Q, Shen H, Chene SM, et al. Synergistic antifungal effect of glabridin and fluconazole. PLoS One 9: e103442.
    Pubmed KoreaMed CrossRef
  32. Shinde RB, Raut JS, Chauhan NM, Karuppayil SM. 2013. Chloroquine sensitizes biofilms of Candida albicans to antifungal azoles. Braz. J. Infect. Dis. 17: 395-400.
    Pubmed CrossRef
  33. Ahmad A, Khan A, Manzoor N, Khan LA. 2010. Evolution of ergosterol biosynthesis inhibitors as fungicidal against Candida. Microb. Pathog. 48: 35-41.
    Pubmed CrossRef
  34. Park HW, Choi KD, Shin IS. 2013. Antimicrobial activity of isothiocyanates (ITCs) extracted from horseradish (Armoracia rusticana) root against oral microorganisms. Biocontrol. Sci. 18: 163-168.
    Pubmed CrossRef
  35. Guiamet PS, Gómez de Saravia SG. 2005. Laboratory studies of biocorrosion control using traditional and environmentally friendly biocides: an overview. Lat. Am. Appl. Res. 35: 295-300.
  36. Ramage G, Rajendran R, Sherry L, Williams C. 2012. Fungal biofilm resistance. Int. J. Microbiol. 2012: 528521.
    Pubmed KoreaMed CrossRef
  37. Verstrepen KJ, Klis FM. 2006. Flocculation, adhesion and biofilm formation in yeasts. Mol. Microbiol. 60: 5-15.
    Pubmed CrossRef
  38. Vila T, Romo JA, Pierce CG, McHardy SF, Saville SP, LopezRibot JL. 2016. Targeting Candida albicans filamentation for antifungal drug development. Virulence 2016: DOI: 10.1080/21505594.2016.1197444.
    CrossRef
  39. Nickerson KW, Atkin AL, Hornby JM. 2006. Quorum sensing in dimorphic fungi: farnesol and beyond. Appl. Environ. Microbiol. 72: 3805-3813.
    Pubmed KoreaMed CrossRef
  40. Cui J, Ren B, Tong Y, Dai H, Zhang L. 2015. Synergistic combinations of antifungals and anti-virulence agents to fight against Candida albicans. Virulence 6: 362-371.
    Pubmed KoreaMed CrossRef
  41. Turgis M, Han J, Caillet S, Lacroix M. 2009. Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi. Food Control 20: 1073-1079.
    CrossRef
  42. Raut JS, Karuppayil SM. 2014. A status review on the medicinal properties of essential oils. Ind. Crops Prod. 62:250-264.
    CrossRef
  43. Xiao D, Srivastava SK, Lew KL, Zeng Y, Hershberger P, Johnson CS, et al. 2003. Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G2/M arrest and inducing apoptosis. Carcinogenesis 24: 891-897.
    Pubmed CrossRef
  44. National Toxicology Program (NTP). 1982. Carcinogenesis bioassay of allyl isothiocyanate (CAS No. 57-06-7) in F344/N Rats and B6C3F1 mice (Gavage study). Natl. Toxicol. Prog. Tech. Rep. Ser. 234: 1-142.
    Pubmed
  45. Zhang Y. 2010. Allyl isothiocyanate as a cancer chemopreventive phytochemical. Mol. Nutr. Food Res. 54: 127-135.
    Pubmed KoreaMed CrossRef