Antimicrobial, Antibiofilm and Immunomodulatory Activities of Lactobacillus rhamnosus and Lactobacillus gasseri against some Bacterial Pathogens


  • Doaa Mohamed Osama Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University
  • Walid Faisal Elkhatib Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University
  • Amany M. Tawfeik Department of Microbiology, Faculty of Medicine (for girls), Al-Azhar University
  • Mohammad Mabrouk Aboulwafa Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University
  • Nadia Abdel-Haleem Hassouna Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University



Probiotics, Lactobacillus rhamnosus, Lactobacillus gasseri, Biofilm, Antibiofilm, Gamma interferon


In this study, two Lactobacillus (LAB) strains namely, Lactobacillus rhamnosus EMCC 1105 (L. rhamnosus) and Lactobacillus gasseri EMCC 1930 (L. gasseri) were tested for their antagonistic activities against Pseudomonas aeruginosa (P. aeruginosa), Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) which are known to be frequently implicated in biofilm infections. The acidic cell free culture supernatant (CFS) of 24 h and 48 h cultures of both LAB stains showed antimicrobial effects against the three pathogens in radial diffusion assay. These effects were abolished upon neutralization of CFS indicating that this effect was due to acids only. Both LAB strains could effectively inhibit the biofilm formation of the three test pathogens and largely replaced them on polystyrene surfaces as demonstrated by crystal violet staining, viable count and scanning electron microscopy. Both of the tested LAB strains could inhibit the protease productivity of S. aureus in 24 h and 48 h dual species-biofilms. The supernatant of 24 h-dual biofilms of P. aeruginosa with L. gasseri also showed a significantly lower protease activity compared to that of P. aeruginosa individual biofilm. Neither LAB strains affected phospholipase C production by the test pathogens when they co-exist during biofilm formation. The different preparations of LAB strains caused no significant change in the levels of gamma interferon expressed by peripheral blood mononuclear cells in response to stimulation by the test pathogens in vitro. In conclusion, L. gasseri and L. rhamnosus can be considered as promising tools for combating biofilm infections.


[1]Williams NT. Probiotics. Am J Health-Syst Pharm AJHP Off J Am Soc Health-Syst Pharm 2010; 67(6): 449-58.
[2]Turpin W, Humblot C, Thomas M, Guyot J-P. Lactobacilli as multifaceted probiotics with poorly disclosed molecular mechanisms. Int J Food Microbiol 2010; 143(3): 87-102.
[3]Czaplewski L, Bax R, Clokie M, Dawson M, Fairhead H, Fischetti VA, et al. Alternatives to antibiotics—a pipeline portfolio review. Lancet Infect Dis 2016; 16(2): 239-51.
[4]Hume ME. Historic perspective: Prebiotics, probiotics, and other alternatives to antibiotics. Poult Sci 2011; 90(11): 2663-9.
[5]WHO | Antimicrobial resistance
[Internet]. WHO.
[cited 2016Aug 28]. Available from: factsheets/fs194/en/
[6]Culligan EP, Hill C, Sleator RD. Probiotics and gastrointestinal disease: successes, problems and future prospects. Gut Pathog 2009; 1: 19.
[7]Al-Ghazzewi FH, Tester RF. Effect of konjac glucomannan hydrolysates and probiotics on the growth of the skin bacterium Propionibacterium acnes in vitro. Int J Cosmet Sci 2010; 32(2): 139-42.
[8]Bowe WP, Logan AC. Acne vulgaris, probiotics and the gut-brain-skin axis - back to the future? Gut Pathog 2011 Jan 31; 3: 1.
[9]Cribby S, Taylor M, Reid G. Vaginal Microbiota and the Use of Probiotics. Interdiscip Perspect Infect Dis
[Internet]. 2008
[cited 2016 Aug 28]; 2008.
[10]Valdéz JC, Peral MC, Rachid M, Santana M, Perdigón G. Interference of Lactobacillus plantarum with Pseudomonas aeruginosa in vitro and in infected burns: the potential use of probiotics in wound treatment. Clin Microbiol Infect Off Publ Eur Soc Clin Microbiol Infect Dis 2005; 11(6): 472-9.
[11]Oelschlaeger TA. Mechanisms of probiotic actions – A review. Int J Med Microbiol 2010; 300(1): 57-62.
[12]Lebeer S, Vanderleyden J, De Keersmaecker SCJ. Genes and molecules of lactobacilli supporting probiotic action. Microbiol Mol Biol Rev MMBR 2008; 72(4): 728-764.
[13]Römling U, Balsalobre C. Biofilm infections, their resilience to therapy and innovative treatment strategies. J Intern Med 2012; 272(6): 541-61.
[14]Vuotto C, Longo F, Donelli G. Probiotics to counteract biofilm-associated infections: promising and conflicting data. Int J Oral Sci 2014; 6(4): 189-94.
[15]Wu H, Moser C, Wang H-Z, Høiby N, Song Z-J. Strategies for combating bacterial biofilm infections. Int J Oral Sci 2015; 7(1): 1-7.
[16]Rumbaugh KP, Ahmad I. Antibiofilm Agents: From Diagnosis to Treatment and Prevention. Springer Science & Business Media 2014; p. 495.
[17]Miquel S, Lagrafeuille R, Souweine B, Forestier C. Anti-biofilm Activity as a Health Issue. Front Microbiol
[Internet]. 2016 Apr 26
[cited 2016 Oct 27]; 7.
[18]Coconnier MH, Liévin V, Bernet-Camard MF, Hudault S, Servin AL. Antibacterial effect of the adhering human Lactobacillus acidophilus strain LB. Antimicrob Agents Chemother 1997; 41(5): 1046-52. Available from:
[19]Zago CE, Silva S, Sanitá PV, Barbugli PA, Dias CMI, Lordello VB, et al. Dynamics of Biofilm Formation and the Interaction between Candida albicans and Methicillin-Susceptible (MSSA) and -Resistant Staphylococcus aureus(MRSA). PLOS One 2015; 10(4): e0123206.
[20]O’Toole GA. Microtiter Dish Biofilm Formation Assay. J Vis Exp
[Internet]. 2011 Jan 30
[cited 2016 Mar 29]; (47). Available from: ID=2437.
[21]Zhang S, Maddox CW. Cytotoxic Activity of Coagulase-Negative Staphylococci in Bovine Mastitis. Infect Immun. 2000; 68(3): 1102-8. Available from:
[22]Nicodème M, Grill J-P, Humbert G, Gaillard J-L. Extracellular protease activity of different Pseudomonas strains: dependence of proteolytic activity on culture conditions. J Appl Microbiol 2005; 99(3): 641-8.
[23]Hafez MM. Studies Concerning Microbial Adherence To Mammalian Cells. Microbiology and Immunology Department. Cairo, Egypt, Ain Shams University. PhD: 419. Department of Microbiology & Immunology, Ain Shams University, Cairo, Egypt 2005.
[24]Elleboudy NS, Aboulwafa MM, Hassouna NA-H. Phospholipase C from Pseudomonas aeruginosa and Bacillus cereus; characterization of catalytic activity. Asian Pac J Trop Med 2014; 7(11): 860-6.
[25]Fischer ER, Hansen BT, Nair V, Hoyt FH, Dorward DW. Scanning Electron Microscopy. In: Coico R, Kowalik T, Quarles J, Stevenson B, Taylor R, editors. Current Protocols in Microbiology
[Internet]. Hoboken, NJ, USA: John Wiley & Sons, Inc.; 2012
[cited 2016 Oct 27]. Available from:
[26]Xaplanteri P, Lagoumintzis G, Dimitracopoulos G, Paliogianni F. Synergistic regulation of Pseudomonas aeruginosa-induced cytokine production in human monocytes by mannose receptor and TLR2. Eur J Immunol 2009; 39(3): 730-40.
[27]Ventola CL. The Antibiotic Resistance Crisis. Pharm Ther 2015; 40(4): 277-83. Available from: http://www.ncbi.nlm.nih. gov/pmc/articles/PMC4378521/
[28]Guidelines for the Evaluation of Probiotics in Food Report of a Joint FAO/WHO Working Group on Drafting Guidelines for the Evaluation of Probiotics in Food
[cited 2016 Sep 1]. Available from: a0512e/a0512e00.pdf
[29]Lin X, Chen X, Chen Y, Jiang W, Chen H. The effect of five probiotic lactobacilli strains on the growth and biofilm formation of Streptococcus mutans. Oral Dis 2015; 21(1): e128-34.
[30]Sharma D, Saharan BS. Functional characterization of biomedical potential of biosurfactant produced by Lactobacillus helveticus. Biotechnol Rep 2016; 11: 27-35.
[31]Bernal P, Llamas MA. Promising biotechnological applications of antibiofilm exopolysaccharides. Microb Biotechnol 2012; 5(6): 670-3.
[32]Ren D, Li C, Qin Y, Yin R, Li X, Tian M, et al. Inhibition of Staphylococcus aureus adherence to Caco-2 cells by lactobacilli and cell surface properties that influence attachment. Anaerobe 2012; 18(5): 508-15.
[33]Dubin G, Koziel J, Pyrc K, Wladyka B, Potempa J. Bacterial proteases in disease - role in intracellular survival, evasion of coagulation/ fibrinolysis innate defenses, toxicoses and viral infections. Curr Pharm Des 2013; 19(6): 1090-113. Available from:
[34]Titball RW. Bacterial phospholipases C. Microbiol Rev 1993; 57(2): 347-66. Available from: pmc/articles/PMC372913/
[35]Shtrichman R, Samuel CE. The role of gamma interferon in antimicrobial immunity. Curr Opin Microbiol 2001; 4(3): 251-9.