Skip to main content

Quantitative Appraisal of the Probiotic Attributes and In Vitro Adhesion Potential of Anti-listerial Bacteriocin-producing Lactic Acid Bacteria

Probiotics and Antimicrobial Proteins volume 5, pages 99–109 (2013)Cite this article

Abstract

Estimation of bile tolerance, endurance to gastric and intestinal environment and adhesion potential to intestinal cells are significant selection criteria for probiotic lactic acid bacteria (LAB). In this paper, the probiotic potential of native bacteriocin-producing LAB isolated previously from indigenous source has been determined through quantitative approaches. Among fifteen anti-listerial bacteriocin-producing native LAB, ten strains were found to be bile tolerant. The presence of bile salt hydrolase (bsh) gene in native Lactobacillus plantarum strains was detected by PCR and confirmed by nucleic acid sequencing of a representative amplicon. Interestingly, three native LAB strains exhibited significant viability in simulated gastric fluid, analogous to the standard LAB Lactobacillus rhamnosus GG, while an overwhelming majority of the native LAB strains demonstrated the ability to survive and remain viable in simulated intestinal fluid. Quantitative adhesion assays based on conventional plating method and a fluorescence-based method revealed that the LAB isolates obtained from dried fish displayed significant in vitro adhesion potential to human adenocarcinoma HT-29 cells, and the adhesion level was comparable to some of the standard probiotic LAB strains. The present study unravels putative probiotic attributes in certain bacteriocin-producing LAB strains of non-human origin, which on further in vivo characterization could find specific applications in probiotic food formulations targeted for health benefits.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3

References

  1. Coolbear T, Crow V, Harnett J, Harvey S, Holland R, Martley F (2008) Developments in cheese microbiology in New Zealand—Use of starter and non-starter lactic acid bacteria and their enzymes in determining flavour. Int Dairy J 18:705–713

    Article  CAS  Google Scholar 

  2. Leroy F, De Vuyst L (2004) Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci Technol 15:67–78

    Article  CAS  Google Scholar 

  3. Jagannath A, Ramesh A, Ramesh MN, Chandrashekar A, Varadaraj MC (2001) Predictive model for the behavior of Listeria monocytogenes Scott A in Shrikhand, prepared with a biopreservative pediocin K7. Food Microbiol 18:335–343

    Article  CAS  Google Scholar 

  4. Jones RJ, Hussein HM, Zagorec M, Brightwell G, Tagg JR (2008) Isolation of lactic acid bacteria with inhibitory activity against pathogens and spoilage organisms associated with fresh meat. Food Microbiol 25:228–234

    Article  CAS  Google Scholar 

  5. Singh AK, Ramesh A (2008) Succession of dominant and antagonistic lactic acid bacteria in fermented cucumber: insights from a PCR-based approach. Food Microbiol 25:278–287

    Article  CAS  Google Scholar 

  6. Singh AK, Mukherjee S, Adhikari MD, Ramesh A (2012) Fluorescence-based comparative evaluation of bactericidal potency and food application potential of anti-listerial bacteriocin produced by lactic acid bacteria isolated from indigenous samples. Probiotics Antimicrob Proteins 4:122–132

    Article  CAS  Google Scholar 

  7. Lebeer S, Vanderleyden J, De Keersmaecker SC (2008) Genes and molecules of lactobacilli supporting probiotic action. Microbiol Mol Biol Rev 72:728–764

    Article  CAS  Google Scholar 

  8. Marco ML, Pavan S, Kleerebezem M (2006) Towards understanding molecular modes of probiotic action. Curr Opin Biotechnol 17:204–210

    Article  CAS  Google Scholar 

  9. Ouwehand AC, Seppo Salminen, Isolauri E (2002) Probiotics: an overview of beneficial effects. Antonie Van Leeuwenhoek 82:279–289

    Article  CAS  Google Scholar 

  10. Servin AL, Coconnier MH (2003) Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens. Best Pract Res Clin Gastroenterol 17:741–754

    Article  CAS  Google Scholar 

  11. Yan F, Polk DB (2010) Probiotics: progress toward novel therapies for intestinal diseases. Curr Opin Gastroenterol 26:95–101

    Article  Google Scholar 

  12. Alander M, Satokari R, Korpela R, Saxelin M, Vilpponen-Salmela T, Mattila-Sandholm T (1999) Persistence of colonization of human colonic mucosa by a probiotic strain, Lactobacillus rhamnosus GG, after oral consumption. Appl Environ Microbiol 65:351–354

    CAS  Google Scholar 

  13. Elliott SN, Buret MW, Miller MJS, Wallace JL (1998) Bacteria rapidly colonize and modulate healing of gastric ulcers in rats. Am J Physiol Gastrointest Liver Physiol 275:425–432

    Google Scholar 

  14. Collado MC, Meriluoto J, Salminen S (2007) Role of commercial probiotic strains against human pathogen adhesion to intestinal mucus. Lett Appl Microbiol 45:454–460

    Article  CAS  Google Scholar 

  15. Reid G, Burton J (2002) Use of Lactobacillus to prevent infection by pathogenic bacteria. Microbes Infect 4:319–324

    Article  Google Scholar 

  16. Pagnini C, Saeed R, Bamias G, Aresneau KO, Pizarro TT, Cominelli F (2010) Probiotics promote gut health through stimulation of epithelial innate immunity. Proc Natl Acad Sci 107:454–459

    Article  CAS  Google Scholar 

  17. Zhou JS, Gill HS (2005) Immunostimulatory probiotic Lactobacillus rhamnosus HN001 and Bifidobacterium lactis HN019 do not induce pathological inflammation in mouse model of experimental autoimmune thyroiditis. Int J Food Microbiol 103:97–104

    Article  CAS  Google Scholar 

  18. Nueno-Palop C, Narbad A (2011) Probiotic assessment of Enterococcus faecalis CP58 isolated from human gut. Int J Food Microbiol 145:390–394

    Article  CAS  Google Scholar 

  19. Charteris WP, Kelly PM, Morelli L, Collins JK (1998) Development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J Appl Microbiol 84:759–768

    Article  CAS  Google Scholar 

  20. Huang Y, Adams MC (2004) In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionibacteria. Int J Food Microbiol 91:253–260

    Article  Google Scholar 

  21. Gopal PK, Prasad J, Smart J, Gill HS (2001) In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobactirium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli. Int J Food Microbiol 67:207–216

    Article  CAS  Google Scholar 

  22. Sambuy Y, De Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F (2005) The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics. Cell Biol Toxicol 21:1–26

    Article  CAS  Google Scholar 

  23. Tuomola EM, Salminen SJ (1998) Adhesion of some probiotic and dairy Lactobacillus strains to Caco-2 cell cultures. Int J Food Microbiol 5:45–51

    Article  Google Scholar 

  24. Forestier C, De Champs C, Vatoux C, Joly B (2001) Probiotic activities of Lactobacillus casei rhamnosus: in vitro adherence to intestinal cells and antimicrobial properties. Res Microbiol 152:167–173

    Article  CAS  Google Scholar 

  25. Binachi MA, Del Rio D, Pellegrini N, Sansebastiano G, Neviani E, Brighenti F (2004) A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells. Lett Appl Microbiol 39:301–305

    Article  Google Scholar 

  26. Blay GL, Fliss I, Lacroix C (2004) Comparative detection of bacterial adhesion to Caco-2 cells with ELISA, radioactivity and plate count method. J Microbiol Methods 59:211–221

    Article  Google Scholar 

  27. Lee YK, Ho PS, Low CS, Arvilommi H, Salminen S (2004) Permanent colonization by Lactobacillus casei is hindered by the low rate of cell division in mouse gut. Appl Environ Microbiol 70:670–674

    Article  CAS  Google Scholar 

  28. Chang JH, Shim YY, Cha SK, Chee KM (2010) Probiotic characteristics of lactic acid bacteria isolated from kimchi. J Appl Microbiol 109:220–230

    Article  CAS  Google Scholar 

  29. Vitali B, Minervini G, Rizzello CG, Spisni E, Maccaferri S, Brigidi P, Gobbetti M, Di Cagno R (2012) Novel probiotic candidates for humans isolated from raw fruits and vegetables. Food Microbiol 31:116–125

    Article  CAS  Google Scholar 

  30. Singh AK, Ramesh A (2009) Evaluation of a facile method of template DNA preparation for PCR-based detection and typing of lactic acid bacteria. Food Microbiol 26:513–540

    Article  Google Scholar 

  31. Jacobsen CN, Rosenfeldt NV, Hayford AE, Møller PL, Michaelsen KF, Paerregaard A, Sandstrom B, Tvede M, Jakobsen M (1999) Screening of probiotic activities of forty-seven strains of Lactobacillus spp. by in vitro techniques and evaluation of the colonization ability of five selected strains in humans. Appl Environ Microbiol 65:4949–4956

    CAS  Google Scholar 

  32. Ohland CL, MacNaughton WK (2010) Probiotic bacteria and intestinal epithelial barrier function. Am J Physiol Gastrointest Liver Physiol 298:807–810

    Article  Google Scholar 

  33. Begley M, Gahan CGM, Hill C (2005) The interaction between bacteria and bile. FEMS Microbiol Rev 29:625–651

    Article  CAS  Google Scholar 

  34. Chateau N, Deschamps AM, Hadj SA (1994) Heterogeneity of bile salts resistance in the Lactobacillus isolates of a probiotic consortium. Lett Appl Microbiol 18:42–44

    Article  Google Scholar 

  35. Burns P, Vinderola G, Binetti A, Quiberoni A, de los Reyes-Gavilan C, Reinheimer J (2008) Bile-resistant derivatives obtained from non-intestinal dairy lactobacilli. Int Dairy J 18:377–385

    Article  CAS  Google Scholar 

  36. Grill JP, Cayuela C, Antoine JM, Schneider F (2000) Isolation and characterization of a Lactobacillus amylovorus mutant depleted in conjugated bile salt hydrolase activity: relation between activity and bile salt resistance. J Appl Microbiol 89:553–563

    Article  CAS  Google Scholar 

  37. Kaushik JK, Kumar A, Duary RK, Mohanty AK, Grover S, Batish VK (2009) Functional and probiotic attributes of an indigenous isolate of Lactobacillus plantarum. PLoS ONE 4:1–11

    Article  Google Scholar 

  38. Lambert JM, Bongers RS, de Vos WM, Kleerebezem M (2008) Functional analysis of four bile salt hydrolase and penicillin acylase family members in Lactobacillus plantarum WCFS1. Appl Environ Microbiol 74:4719–4726

    Article  CAS  Google Scholar 

  39. Jones BV, Begley M, Hill C, Gahan CGM, Marchesi JR (2008) Functional and comparative metagenomic analysis of bile salt hydrolase activity in the human gut microbiome. Proc Natl Acad Sci 105:13580–13585

    Article  CAS  Google Scholar 

  40. McAuliffe O, Cano RJ, Klaenhammer TR (2005) Genetic analysis of two bile salt hydrolase activities in Lactobacillus acidophilus NCFM. Appl Environ Microbiol 71:4925–4929

    Article  CAS  Google Scholar 

  41. Patel AK, Singhania RR, Pandey A, Chincholkar SB (2010) Probiotic bile salt hydrolase: current developments and perspectives. Appl Biochem Biotechnol 162:166–180

    Article  CAS  Google Scholar 

  42. Liong MT, Shah NP (2005) Acid and bile tolerance and cholesterol removal ability of lactobacilli strains. J Dairy Sci 88:55–66

    Article  CAS  Google Scholar 

  43. Blum S, Haller D, Pfeifer A, Schiffrin EJ (2002) Probiotics and immune response. Clin Rev Allergy Immunol 22:287–309

    Article  Google Scholar 

  44. Langerholc T, Maragkoudakis PA, Wollgast J, Gradisnik L, Cencic A (2011) Novel and established cell line models—An indispensable tool in food science and nutrition. Trends Food Sci Technol 22:S11–S20

    Article  CAS  Google Scholar 

  45. Ossowski I, Reunanen J, Satokari R, Vesterlund S, Kankainen M, Huhtinen H, Tynkkynen S, Salminen S, De Vos WM, Palva A (2010) Mucosal adhesion properties of the probiotic Lactobacillus rhamnosus GG SpaCBA and SpaFED pilin subunits. Appl Environ Microbiol 76:2049–2057

    Article  Google Scholar 

  46. Jensen H, Grimmer S, Naterstad K, Axelsson L (2012) In vitro testing of commercial and potential probiotic lactic acid bacteria. Int J Food Microbiol 153:216–222

    Article  Google Scholar 

  47. Tallon R, Arias S, Bressollier P, Urdaci MC (2007) Strain- and matrix-dependent adhesion of Lactobacillus plantarum is mediated by proteinaceous bacterial compounds. J Appl Microbiol 102:442–451

    Article  CAS  Google Scholar 

  48. Archimbaud C, Shankar N, Forestier C, Baghdayan A, Gilmore MS, Charbonne F, Joly B (2002) In vitro adhesive properties and virulence factors of Enterococcus faecalis strains. Res Microbiol 153:75–80

    Article  CAS  Google Scholar 

  49. Hoefel D, Groobya WL, Monisa PT, Andrews S, Saint CP (2003) A comparative study of carboxyfluorescein diacetate and carboxyfluorescein diacetate succinimidyl ester as indicators of bacterial activity. J Microbiol Methods 52:379–388

    Article  CAS  Google Scholar 

  50. Ramiah K, van Reenen CA, Dicks LMT (2008) Surface-bound proteins of Lactobacillus plantarum 423 that contribute to adhesion of Caco-2 cells and their role in competitive exclusion and displacement of Clostridium sporogenes and Enterococcus faecalis. Res Microbiol 159:470–475

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a research grant from Council of Scientific and Industrial Research (CSIR), New Delhi, Government of India [No. 38(1251)/10/EMR-II]. We thank the National Facility of Automated DNA Sequencing, Department of Biochemistry, Delhi University, South Campus for their support in nucleic acid sequencing.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Affiliations

  1. Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, 781 039, Assam, India

    Sandipan Mukherjee, Atul Kumar Singh, Manab Deb Adhikari & Aiyagari Ramesh

Authors
  1. Sandipan Mukherjee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Atul Kumar Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manab Deb Adhikari
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Aiyagari Ramesh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Aiyagari Ramesh.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Mukherjee, S., Singh, A.K., Adhikari, M.D. et al. Quantitative Appraisal of the Probiotic Attributes and In Vitro Adhesion Potential of Anti-listerial Bacteriocin-producing Lactic Acid Bacteria. Probiotics & Antimicro. Prot. 5, 99–109 (2013). https://doi.org/10.1007/s12602-013-9131-5

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12602-013-9131-5

Keywords

  • Lactic acid bacteria
  • Probiotic
  • Adhesion
  • Bacteriocin