Parinaz GHADAM, Rouha KASRA KERMANSHAHI, Roya VAHEDI SHAHANDASHTI

The inhibitory effect of bacteriocin produced by Lactobacillus acidophilus ATCC 4356 and Lactobacillus plantarum ATCC 8014 on planktonic cells and biofilms of Serratia marcescens

Background/aim: The spread of antibiotic-resistant pathogens has resulted in the need for new treatments. The aim of the present study is to investigate the effect of bacteriocin from Lactobacillus acidophilus ATCC 4356 and Lactobacillus plantarum ATCC 8014 on planktonic and biofilm forms of Serratia marcescens strains. Materials and methods: The direct antagonism of the L. plantarum and L. acidophilus cell-free supernatant on S. marcescens cultures was determined using an optical density assay. The bacteriocin was partial purified by ammonium sulfate precipitation. Its molecular weight was analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The effect of bacteriocins on the biofilm of S. marcescens strains was then determined with 2,3,5-triphenyl tetrazolium chloride. Results: The purified bacteriocin from L. plantarum ATCC 8014 and partially purified bacteriocin from L. acidophilus ATCC 4356 displayed noticeable inhibitory activity against planktonic and biofilm forms of S. marcescens strains. SDS-PAGE analysis revealed that the apparent molecular weight of bacteriocin from L. planetarium was 63 kDa, and that of bacteriocin from L. acidophilus was 68 or 48 kDa. Conclusion: The bacteriocins could be effective compounds to control surface-attached pathogenic bacteria and can be used as therapeutic agents after acceptable in vivo experimentation.

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1. Costerton JW. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284: 1318-1322.
2. Anwar H, Dasgupta MK, Costerton JW. Testing the susceptibility of bacteria in biofilms to antibacterial agents. Antimicrob Agents Chemother 1990; 34: 2043-2046.
3. Campanac C, Pineau L, Payard A, Roques C. Interactions between biocide cationic agents and bacterial biofilms. Antimicrob Agents Chemother 2002; 46: 1469-1474.
4. Drenkard E. Antimicrobial resistance of Pseudomonas aeruginosa biofilms. Microbes Infect 2003; 5: 1213-1219.
5. Costerton JW, Cheng KJ, Geesey GG, Ladd TI, Nickel JC, Dasgupta M, Marrie TJ. Bacterial biofilms in nature and disease. Ann Rev Microbiol 1987; 41: 435-464.
6. Parsek MR, Singh PK. Bacterial biofilms: an emerging link to disease pathogenesis. Ann Rev Microbiol 2003; 57: 677-701.
7. Pitts B, Hamilton MA, Zelver N, Stewart PS. A microtiterplate screening method for biofilm disinfection and removal. J Microbiol Methods 2003; 54: 269-276.
8. Hejazi A, Falkiner FR (1997). Serratia marcescens. J Med Microbiol 1997; 46: 903-912.
9. Slater H, Crow M, Everson L, Salmond GPC. Phosphate availability regulates biosynthesis of two antibiotics, prodigiosin and carbapenem, in Serratia via both quorumsensing-dependent and -independent pathways. Mol Microbiol 2003; 47: 303-320.
10. Anderson GG, Toole GAO. Innate and induced resistance mechanisms of bacterial biofilms. Curr Top Microbiol Immunol 2008; 322: 85-105.
11. Cotter PD, Ross RP, Hill C. Bacteriocins - a viable alternative to antibiotics? Nat Rev Microbiolm 2013; 11: 95-105.
12. Cleveland J, Montville TJ, Nes IF, Chikindas ML. Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 2001; 71: 1-20.
13. Lash BW, Mysliwiec TH, Gourama H. Detection and partial characterization of a broad-range bacteriocin produced by Lactobacillus plantarum (ATCC 8014). Food Microbiol 2005; 22: 199-204.
14. Oh S, Kim SH, Worobo RW. Characterization and purification of a bacteriocin produced by a potential probiotic culture, Lactobacillus acidophilus 30SC. J Dairy Sci 2000; 83: 2747- 2752.
15. Lash BW, Gourama H. Microscale assay for screening of inhibitory activity of Lactobacillus. BioTechniques 2002; 33: 1224-1228.
16. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K, editors. Short Protocols in Molecular Biology. 4th ed. New York, USA: John Wiley & Sons, Inc.; 1999.
17. Zor T, Selinger Z. Linearization of the Bradford protein assay increases its sensitivity: theoretical and experimental studies. Anal Biochem 1996; 236: 302-308.
18. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685.
19. Giacometti A, Cirioni O, Del Prete MS, Barchiesi F, Paggi AM, Petrelli E, Scalise G. Comparative activities of polycationic peptides and clinically used antimicrobial agents against multidrug-resistant nosocomial isolates of Acinetobacter baumannii. J Antimicrob Chemother 2000; 46: 807-810.
20. Nielsen DS, Cho G, Hanak A, Huch M, Franz C, Arneborg N. The effect of bacteriocin-producing Lactobacillus plantarum strains on the intracellular pH of sessile and planktonic Listeria monocytogenes single cells. Int J Food Microbiol 2010; 141: S53-S59.
21. Okuda K, Zendo T, Sugimoto S, Iwase T, Tajima A, Yamada S, Sonomoto K, Mizunoe Y. Effects of bacteriocins on methicillinresistant Staphylococcus aureus biofilm. Antimicrob Agents Chemother 2013; 57: 5572-5579.
22. Knezevic P, Petrovic O. A colorimetric microtiter plate method for assessment of phage effect on Pseudomonas aeruginosa biofilm. J Microbiol Methods 2008; 74: 114-118.
23. Labbate M, Queck SY, Koh KS, Rice SA, Givskov M, Kjelleberg S. Quorum sensing-controlled biofilm development in Serratia liquefaciens MG1. J Bacteriol 2004; 186: 692-698.
24. Shakeri S, Kermanshahi RK, Moghaddam MM, Emtiazi G. Assessment of biofilm cell removal and killing and biocide efficacy using the microtiter plate test. Biofouling 2007; 23: 79- 86.
25. Shanks RMQ, Dashiff A, Alster JS, Kadouri DE. Isolation and identification of a bacteriocin with antibacterial and antibiofilm activity from Citrobacter freundii. Arch Microbiol 2012; 194: 575-587.
26. Adukwu EC, Allen SCH, Phillips CA. The anti-biofilm activity of lemongrass (Cymbopogon flexuosus) and grapefruit (Citrus paradisi) essential oils against five strains of Staphylococcus aureus. J Appl Microbiol 2012; 113: 1217-1227.
27. Stewart PS. Mechanisms of antibiotic resistance in bacterial biofilms. Int J Med Microbiol 2002; 292: 107-113. 28. Fuller R. Probiotics in man and animals. J Appl Bacteriol 1989; 66: 365-378.
29. Stevens KA, Sheldon BW, Klapes NA, Klaenhammer TR. Nisin treatment for inactivation of Salmonella species and other gram-negative bacteria. Appl Environ Microbiol 1991; 57: 3613-3615.
30. Aroutcheva A, Gariti D, Simon M, Shott S, Faro J, Simoes JA, Gurguis A, Faro S. Defense factors of vaginal lactobacilli. Am J Obstet Gynecol 2001; 185: 375-379.
31. Riaz S, Nawaz SK, Hasnain SH. Bacteriocins produced by L. fermentum and L. acidophilus can inhibit cephalosporin resistant E. coli. Braz J Microbiol 2010; 41: 643-648.
32. Smaoui S, Elleuch L, Bejar W, Karray-Rebai I, Ayadi I, Jaouadi B, Mathieu F, Chouayekh H, Bejar S, Mellouli L. Inhibition of fungi and Gram-negative bacteria by bacteriocin BacTN635 produced by Lactobacillus plantarum sp. TN635. Appl Biochem Biotechnol 2010; 162: 1132-1146.
33. Tahmourespour A, Kermanshahi RK. The effect of a probiotic strain (Lactobacillus acidophilus) on the plaque formation of oral Streptococci. Bosn J Basic Med Sci 2011; 11: 37-40.
34. Vuotto C, Longo F, Donelli G. Probiotics to counteract biofilmassociated infections: promising and conflicting data. Int J Oral Sci 2014; 6: 189-194.
35. Aslim B, Yuksekdag ZN, Sarikaya E, Beyatli Y. Determination of the bacteriocin-like substances produced by some lactic acid bacteria isolated from Turkish dairy products. LWT-Food Sci Technol 2005; 38: 691-694.
36. Lauková A, Mareková M, Javorský P. Detection and antimicrobial spectrum of a bacteriocin‐like substance produced by Enterococcus faecium CCM4231. Lett Appl Microbiol 1993; 16: 257-260.
37. Arihara K, Ogihara S, Mukai T, Itoh M, Kondo Y. Salivacin 140, a novel bacteriocin from Lactobacillus salivarius subsp. salicinius T140 active against pathogenic bacteria. Lett Appl Microbiol 1996; 22: 420-424.
38. Messi P, Bondi M, Sabia C, Battini R, Manicardi G. Detection and preliminary characterization of a bacteriocin (plantaricin 35d) produced by a Lactobacillus plantarum strain. Int J Food Microbiol 2001; 64: 193-198.
39. Sauer K, Camper AK, Ehrlich GD, William J, Davies DG, Costerton JW. Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J Bacteriol 2002; 184: 1140-1154.
40. Rice SA, Koh KS, Quick SY, Labbate M, Lam KW, Kjelleberg S. Biofilm formation and sloughing in Serratia marcescens are controlled by quorum sensing and nutrient cues. J Bacteriol 205; 187: 3477-3485.
41. Ramos AN, Gobbato N, Rachid M, González L, Yantorno O, Valdez JC1. Effect of Lactobacillus plantarum and Pseudomonas aeruginosa culture supernatants on polymorphonuclear damage and inflammatory response. Comp Immunol Microbiol Infect Dis 2010; 33: 267-275.