Abstract
Escherichia coli (E. coli) is associated with an array of health-threatening contaminations, some of which are related to biofilm states. The pgaABCD-encoded poly-beta-1,6-N-acetyl-D-glucosamine (PGA) polymer plays an important role in biofilm formation. This study was conducted to determine the inhibitory effect of gallic acid (GA) against E. coli biofilm formation. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of GA against planktonic E. coli were 0.5 and 4 mg/mL, and minimal biofilm inhibitory concentration and minimal biofilm eradication concentration values of GA against E. coli in biofilms were 2 and 8 mg/mL, respectively. Quantitative crystal violet staining of biofilms and ESEM images clearly indicate that GA effectively, dose-dependently inhibited biofilm formation. CFU counting and confocal laser scanning microscopy measurements showed that GA significantly reduced viable bacteria in the biofilm. The contents of polysaccharide slime, protein, and DNA in the E. coli biofilm also decreased. qRT-PCR data showed that at the sub-MIC level of GA (0.25 mg/mL) and expression of pgaABC genes was downregulated, while pgaD gene expression was upregulated. The sub-MBC level of GA (2 mg/mL) significantly suppressed the pgaABCD genes. Our results altogether demonstrate that GA inhibited viable bacteria and E. coli biofilm formation, marking a novel approach to the prevention and treatment of biofilm-related infections in the food industry.
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References
Boehm A, Steiner S, Zaehringer F, Casanova A, Hamburger F, Ritz D, Keck W, Ackermann M, Schirmer T, Jenal U (2009) Second messenger signalling governs Escherichia coli biofilm induction upon ribosomal stress. Mol Microbiol 72(6):1500–1516. https://doi.org/10.1111/j.1365-2958.2009.06739.x
Borges A, Saavedra MJ, Simões M (2012) The activity of ferulic and gallic acids in biofilm prevention and control of pathogenic bacteria. Biofouling 28(7):755–767. https://doi.org/10.1080/08927014.2012.706751
Cerca N, Jefferson KK (2008) Effect of growth conditions on poly- N -acetylglucosamine expression and biofilm formation in Escherichia coli. FEMS Microbiol Lett 283(1):36–41. https://doi.org/10.1111/j.1574-6968.2008.01142.x
Chung KT, Wong TY, Wei CI, Huang YW, Lin Y (1998) Tannins and human health: a review. Crit Rev Food Sci Nutr 38(6):421–464. https://doi.org/10.1080/10408699891274273
Coughlan LM, Cotter PD, Hill C, Alvarezordóñez A (2016) New weapons to fight old enemies: novel strategies for the (bio)control of bacterial biofilms in the food industry. Front Microbiol 7(e26278):1641. https://doi.org/10.3389/fmicb.2016.01641
Cui H, Ma C, Lin L (2016) Synergetic antibacterial efficacy of cold nitrogen plasma and clove oil against Escherichia coli O157:H7 biofilms on lettuce. Food Control 66:8–16. https://doi.org/10.1016/j.foodcont.2016.01.035
Cui H, Yuan L, Li W, Lin L (2017) Edible film incorporated with chitosan and Artemisia annua oil nanoliposomes for inactivation of Escherichia coli O157:H7 on cherry tomato. Int J Food Sci Technol 52(3):687–698. https://doi.org/10.1111/ijfs.13322
Díaz-Gómez R, López-Solís R, Obreque-Slier E, Toledo-Araya H (2013) Comparative antibacterial effect of gallic acid and catechin against Helicobacter pylori. LWT Food Sci Technol 54(2):331–335. https://doi.org/10.1016/j.lwt.2013.07.012
Doucet FJ, Lead JR, Maguire L, Achterberg EP, Millward GE (2005) Visualisation of natural aquatic colloids and particles—a comparison of conventional high vacuum and environmental scanning electron microscopy. J Environ Monit 7(2):115–121. https://doi.org/10.1039/b413832e
Duffy LL, Grau FH, Vanderlinde PB (2000) Acid resistance of enterohaemorrhagic and generic Escherichia coli associated with foodborne disease and meat. Int J Food Microbiol 60(1):83–89. https://doi.org/10.1016/S0168-1605(00)00353-6
Dynes JJ, Lawrence JR, Korber DR, Swerhone GD, Leppard GG, Hitchcock AP (2009) Morphological and biochemical changes in Pseudomonas fluorescens biofilms induced by sub-inhibitory exposure to antimicrobial agents. Can J Microbiol 55(2):163–178. https://doi.org/10.1139/w08-109
Flemming HC, Wingender J (2010) The biofilm matrix. Nat Rev Microbiol 8(9):623–633. https://doi.org/10.1038/nrmicro2415
Høiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O (2010) Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents 35(4):322–332. https://doi.org/10.1016/j.ijantimicag.2009.12.011
Itoh Y, Rice JD, Goller C, Pannuri A, Taylor J, Meisner J, Beveridge TJ, Preston JF 3rd, Romeo T (2008) Roles of pgaABCD genes in synthesis, modification, and export of the Escherichia coli biofilm adhesin poly-beta-1,6-N-acetyl-D-glucosamine. J Bacteriol 190(10):3670–3680. https://doi.org/10.1128/JB.01920-07
Jagani S, Chelikani R, Kim DS (2009) Effects of phenol and natural phenolic compounds on biofilm formation by Pseudomonas aeruginosa. Biofouling 25(4):321–324. https://doi.org/10.1080/08927010802660854
Kulshrestha S, Khan S, Hasan S, Khan ME, Misba L, Khan AU (2016) Calcium fluoride nanoparticles induced suppression of Streptococcus mutans biofilm: an in vitro and in vivo approach. Appl Microbiol Biotechnol 100(4):1901–1914. https://doi.org/10.1007/s00253-015-7154-4
Lee JH, Kim YG, Cho HS, Ryu SY, Cho MH, Lee J (2014a) Coumarins reduce biofilm formation and the virulence of Escherichia coli O157:H7. Phytomedicine 21(8–9):1037–1042. https://doi.org/10.1016/j.phymed.2014.04.008
Lee JH, Kim YG, Ryu SY, Cho MH, Lee J (2014b) Ginkgolic acids and Ginkgo biloba extract inhibit Escherichia coli O157:H7 and Staphylococcus aureus biofilm formation. Int J Food Microbiol 174:47–55. https://doi.org/10.1016/j.ijfoodmicro.2013.12.030
Liao H, Zhang F, Liao X, Hu X, Chen Y, Deng L (2010) Analysis of Escherichia coli cell damage induced by HPCD using microscopies and fluorescent staining. Int J Food Microbiol 144(1):169–176. https://doi.org/10.1016/j.ijfoodmicro.2010.09.017
Limoli DH, Jones CJ, Wozniak DJ (2015) Bacterial extracellular polysaccharides in biofilm formation and function. Microbiol Spectr 3(3). https://doi.org/10.1128/microbiolspec.MB-0011-2014
Limpisophon K, Schleining G (2017) Use of gallic acid to enhance the antioxidant and mechanical properties of active fish gelatin film. J Food Sci 82(1):80–89. https://doi.org/10.1111/1750-3841.13578
Liu KY, Hu S, Chan BC, Wat EC, Lau CB, Hon KL, Fung KP, Leung PC, Hui PC, Lam CW, Wong CK (2013) Anti-inflammatory and anti-allergic activities of Pentaherb formula, Moutan Cortex (Danpi) and gallic acid. Molecules 18(3):2483–2500. https://doi.org/10.3390/molecules18032483
Liu M, Wu X, Li J, Liu L, Zhang R, Shao D, Du X (2017) The specific anti-biofilm effect of gallic acid on Staphylococcus aureus by regulating the expression of the ica operon. Food Control 73:613–618. https://doi.org/10.1016/j.foodcont.2016.09.015
Negi PS (2012) Plant extracts for the control of bacterial growth: efficacy, stability and safety issues for food application. Int J Food Microbiol 156(1):7–17. https://doi.org/10.1016/j.ijfoodmicro.2012.03.006
Newell DG, Koopmans M, Verhoef L, Duizer E, Aidara-Kane A, Sprong H, Opsteegh M, Langelaar M, Threfall J, Scheutz F, van der Giessen J, Kruse H (2010) Food-borne diseases—the challenges of 20 years ago still persist while new ones continue to emerge. Int J Food Microbiol 139(Suppl 1):S3–S15. https://doi.org/10.1016/j.ijfoodmicro.2010.01.021
Oliveira KÁRD, Sousa JPD, Medeiros JADC, Magnani M, Júnior JPDS, Souza ELD (2015) Synergistic inhibition of bacteria associated with minimally processed vegetables in mixed culture by carvacrol and 1,8-cineole. Food Control 47:334–339. https://doi.org/10.1016/j.foodcont.2014.07.014
Prakash B, Veeregowda BM, Krishnappa G (2003) Biofilms: a survival strategy of bacteria. Curr Sci India 85(9):1299–1307
Raffaella C, Casettari L, Fagioli L, Cespi M, Bonacucina G, Baffone W (2017) Activity of essential oil-based microemulsions against Staphylococcus aureus biofilms developed on stainless steel surface in different culture media and growth conditions. Int J Food Microbiol 241:132–140. https://doi.org/10.1016/j.ijfoodmicro.2016.10.021
Ryu JH, Beuchat LR (2005) Biofilm formation by Escherichia coli O157:H7 on stainless steel: effect of exopolysaccharide and Curli production on its resistance to chlorine. Appl Environ Microbiol 71(1):247–254. https://doi.org/10.1128/AEM.71.1.247-254.2005
Serra DO, Mika F, Richter AM, Hengge R (2016) The green tea polyphenol EGCG inhibits E. coli biofilm formation by impairing amyloid curli fibre assembly and downregulating the biofilm regulator CsgD via the sigma(E)-dependent sRNA RybB. Mol Microbiol 101(1):136–151. https://doi.org/10.1111/mmi.13379
Shao D, Li J, Li J, Tang R, Liu L, Shi J, Huang Q, Yang H (2015) Inhibition of gallic acid on the growth and biofilm formation of Escherichia coli and Streptococcus mutans. J Food Sci 80(6):M1299–M1305. https://doi.org/10.1111/1750-3841.12902
Sharma G, Sharma S, Sharma P, Chandola D, Dang S, Gupta S, Gabrani R (2016) Escherichia coli biofilm: development and therapeutic strategies. J Appl Microbiol 121(2):309–319. https://doi.org/10.1111/jam.13078
Sivaranjani M, Prakash M, Gowrishankar S, Rathna J, Pandian SK, Ravi AV (2017) In vitro activity of alpha-mangostin in killing and eradicating Staphylococcus epidermidis RP62A biofilms. Appl Microbiol Biotechnol 101(8):3349–3359. https://doi.org/10.1007/s00253-017-8231-7
Sun X, Wang Z, Kadouh H, Zhou K (2014) The antimicrobial, mechanical, physical and structural properties of chitosan–gallic acid films. LWT Food Sci Technol 57(1):83–89. https://doi.org/10.1016/j.lwt.2013.11.037
Wang X, Preston JF, Romeo T (2004) The pgaABCD locus of Escherichia coli promotes the synthesis of a polysaccharide adhesin required for biofilm formation. J Bacteriol 186(9):2724–2734. https://doi.org/10.1128/JB.186.9.2724-2734.2004
Webber B, Canova R, Esper LM, Perdoncini G, Nascimento VPD, Pilotto F, Santos LRD, Rodrigues LB (2015) The use of vortex and ultrasound techniques for the in vitro removal of Salmonella spp. biofilms. Acta Sci Vet 43(1):1–5
Wood TK, González Barrios AF, Herzberg M, Lee J (2006) Motility influences biofilm architecture in Escherichia coli. Appl Microbiol Biotechnol 72(2):361–367. https://doi.org/10.1007/s00253-005-0263-8
Yoon CH, Chung SJ, Lee SW, Park YB, Lee SK, Park MC (2013) Gallic acid, a natural polyphenolic acid, induces apoptosis and inhibits proinflammatory gene expressions in rheumatoid arthritis fibroblast-like synoviocytes. Joint Bone Spine 80(3):274–279. https://doi.org/10.1016/j.jbspin.2012.08.010
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The authors acknowledge the financial support from National Natural Science Foundation of China (grant no. 31301472) and Project Funded by Fundamental Research Funds for the Central Universities (no. 3102016QD075).
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Kang, J., Li, Q., Liu, L. et al. The specific effect of gallic acid on Escherichia coli biofilm formation by regulating pgaABCD genes expression. Appl Microbiol Biotechnol 102, 1837–1846 (2018). https://doi.org/10.1007/s00253-017-8709-3
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DOI: https://doi.org/10.1007/s00253-017-8709-3