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In Vitro Comparative Analysis of Probiotic and Functional Attributes of Indigenous Isolates of Bifidobacteria

  • Sajan C. Achi
  • Prakash M. HalamiEmail author
Article
  • 48 Downloads

Abstract

In the present study, probiotic, safety and functional characteristics of eight indigenous bifidobacterial isolates were compared to identify suitable strains for functional food application. Among the isolates, six strains of Bifidobacterium longum and one each of Bifidobacterium breve and Bifidobacterium bifidum were identified by 16S rRNA, xfp and hsp60 gene sequencing. Diversity among these strains was established by RAPD and Rep-PCR. Genes associated with sortase-dependent pili (SDP) (credited for role in adhesion) and serpin (immunomodulation) which can serve as potential marker genes for rapid identification of probiotic Bifidobacterium, was also evaluated. All the isolates exhibited potential probiotic, functional (antimicrobial activity, antioxidant activity, phytase activity, milk fermentation ability) and safety attributes. However, among them, B. breve NCIM5671 exhibited, better tolerance to low pH, amylase activity and exopolysaccharide producing ability. B. bifidum NCIM5697 and B. longum NCIM5672 demonstrated higher adherence ability to Caco-2 cells. NCIM5697 also displayed exopolysaccharide producing ability while NCIM5672 showed strong antibacterial activity against pathogens tested. Further, with respect to presence of adhesion marker genes, disparity was observed among B. longum strains. B. longum NCIM5684 and B. longum NCIM5686 displayed presence of subunits of SDP reported to be present in B. breve. In addition, B. longum NCIM5686 also lacked SDP present in all other B. longum isolates. B. breve NCIM5671, B. longum NCIM5672 and B. bifidum NCIM5697 with appreciable traits qualifies as potential probiotic cultures. Further, the variations observed in molecular and functional characteristics of isolates signify genetic diversity among the cultures.

Notes

Acknowledgements

The authors are thankful to The Director CSIR-Central Food Technological Research Institute (CFTRI), Mysuru, India, for providing necessary funds and facilities. SA would like to acknowledge CSIR for Granting Senior Research Fellowship, Dr. Navneet K, for support during cell adhesion studies. This work was supported by Council of Scientific and Industrial Research (CSIR), New Delhi under XIIth five-year plan project (BSC0202).

Funding

This work was funded by Council of Scientific and Industrial Research(CSIR), New Delhi under XIIth five-year plan project (BSC0202).

Compliance with Ethical Standards

Conflict of interest

Authors declare that they have no conflict of interest.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Institute.

Informed Consent

Informed consent was obtained from parents of all infants included in the study.

Supplementary material

284_2018_1615_MOESM1_ESM.doc (850 kb)
Supplementary material 1 (DOC 850 KB)

References

  1. 1.
    Arboleya S, Ruas-Madiedo P, Margolles A, Solís G, Salminen S, Clara G, Gueimonde M (2011) Characterization and in vitro properties of potentially probiotic Bifidobacterium strains isolated from breast-milk. Int J Food Microbiol 149:28–36.  https://doi.org/10.1016/j.ijfoodmicro.2010.10.036 CrossRefPubMedGoogle Scholar
  2. 2.
    Arboleya S, Watkins C, Stanton C, Ross RP (2016) Gut bifidobacteria populations in human health and aging. Front Microbiol.  https://doi.org/10.3389/fmicb.2016.01204 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Archer AC, Halami PM (2015) Probiotic attributes of Lactobacillus fermentum isolated from human feces and dairy products. Appl Microbiol Biotechnol 99:8113–8123.  https://doi.org/10.1007/s00253-015-6679-x CrossRefPubMedGoogle Scholar
  4. 4.
    Baffoni L, Stenico V, Strahsburger E, Gaggìa F, Di Gioia D, Modesto M, Mattarelli P, Biavati B (2013) Identification of species belonging to the Bifidobacterium genus by PCR-RFLP analysis of a hsp60 gene fragment. BMC Microbiol.  https://doi.org/10.1186/1471-2180-13-149 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Bottacini F, Motherway MOC, Kuczynski J, O’Connell KJ, Serafini F, Duranti S, Milani C, Turroni F, Lugli GA, Zomer A (2014) Comparative genomics of the Bifidobacterium breve taxon. BMC Genomics 15:170.  https://doi.org/10.1186/1471-2164-15-170 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Bover-Cid S, Holzapfel WH (1999) Improved screening procedure for biogenic amine production by lactic acid bacteria. Int J Food Microbiol 53:33–41.  https://doi.org/10.1016/S0168-1605(99)00152-X CrossRefPubMedGoogle Scholar
  7. 7.
    Chaplin AV, Efimov BA, Smeianov VV, Kafarskaia LI, Pikina AP, Shkoporov AN (2015) Intraspecies genomic diversity and long-term persistence of Bifidobacterium longum. PLoS ONE.  https://doi.org/10.1371/journal.pone.0135658 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Devi SM, Archer AC, Halami PM (2015) Screening, characterization and in vitro evaluation of probiotic properties among lactic acid bacteria through comparative analysis. Probiotics Antimicrob Proteins 7:181–192.  https://doi.org/10.1007/s12602-015-9195-5 CrossRefPubMedGoogle Scholar
  9. 9.
    European Food Safety Authority (EFSA) (2007) Introduction of a qualified presumption of safety (QPS) approach for assessment of selected microorganisms referred to EFSA. Opinion of the scientific committee (question no EFSA-Q-2005-293). EFSA J 587:1–16.  https://doi.org/10.2903/j.efsa.2007.587 CrossRefGoogle Scholar
  10. 10.
    EFSA BIOHAZ Panel (EFSA Panel on Biological Hazards), Ricci A, Allende A, Bolton D, Chemaly M, Davies R, Girones R, Koutsoumanis K, Herman L, Lindqvist R, Nørrung B, Robertson L, Ru G, Sanaa M, Simmons M, Skandamis P, Snary E, Speybroeck N, Ter Kuile B, Threlfall J, Wahlström H, Cocconcelli PS, Klein G, Peixe L, Maradona MP, Querol A, Suarez JE, Sundh I, Vlak J, Correia S, Fernández Escámez PS (2017) Statement on the update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA 5: suitability of taxonomic units notified to EFSA until September 2016. EFSA J.  https://doi.org/10.2903/j.efsa.2017.4663 CrossRefGoogle Scholar
  11. 11.
    FAO/WHO (2002) WHO working group report on drafting guidelines for the evaluation of probiotics in food. FAO/WHO, LondonGoogle Scholar
  12. 12.
    Foroni E, Serafini F, Amidani D, Turroni F, He F, Bottacini F, Motherway MOC, Viappiani A, Zhang Z, Rivetti C (2011) Genetic analysis and morphological identification of pilus-like structures in members of the genus Bifidobacterium. Microb Cell Fact.  https://doi.org/10.1186/1475-2859-10-S1-S16 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Gevers D, Huys G, Swings J (2001) Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiol Lett 205:31–36.  https://doi.org/10.1111/j.1574-6968.2001.tb10921.x CrossRefPubMedGoogle Scholar
  14. 14.
    Hongpattarakere T, Cherntong N, Wichienchot S, Kolida S, Rastall RA (2012) In vitro prebiotic evaluation of exopolysaccharides produced by marine isolated lactic acid bacteria. Carbohydr Polym 87:846–852.  https://doi.org/10.1016/j.carbpol.2011.08.085 CrossRefGoogle Scholar
  15. 15.
    Jarocki P, Podleśny M, Komoń-Janczara E, Kucharska J, Glibowska A, Targoński Z (2016) Comparison of various molecular methods for rapid differentiation of intestinal bifidobacteria at the species, subspecies and strain level. BMC Microbiol 16:159.  https://doi.org/10.1186/s12866-016-0779-3 CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Křížová J, Španová A, Rittich B (2008) RAPD and rep-PCR fingerprinting for characterization of Bifidobacterium species. Folia Microbiol.  https://doi.org/10.1007/s12223-008-0014-1 CrossRefGoogle Scholar
  17. 17.
    Lee JH, O’Sullivan DJ (2010) Genomic insights into bifidobacteria. Microbiol Mol Biol Rev 74:378–416.  https://doi.org/10.1128/MMBR.00004-10 CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Munoz-Quezada S, Chenoll E, Vieites JM, Genovés S, Maldonado J, Bermúdez-Brito M, Gomez-Llorente C, Matencio E, Bernal MJ, Romero F (2013) Isolation, identification and characterisation of three novel probiotic strains (Lactobacillus paracasei CNCM I-4034, Bifidobacterium breve CNCM I-4035 and Lactobacillus rhamnosus CNCM I-4036) from the faeces of exclusively breast-fed infants. Br J Nutr 109:S51–S62.  https://doi.org/10.1017/S0007114512005211 CrossRefPubMedGoogle Scholar
  19. 19.
    Papadimitriou K, Zoumpopoulou G, Foligné B, Alexandraki V, Kazou M, Pot B, Tsakalidou E (2015) Discovering probiotic microorganisms: in vitro, in vivo, genetic and omics approaches. Front Microbiol 6:58.  https://doi.org/10.3389/fmicb.2015.00058 CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Raghavendra P, Halami PM (2009) Screening, selection and characterization of phytic acid degrading lactic acid bacteria from chicken intestine. Int J Food Microbiol 133:129–134.  https://doi.org/10.1016/j.ijfoodmicro.2009.05.006 CrossRefPubMedGoogle Scholar
  21. 21.
    Rodríguez E, Arqués JL, Rodríguez R, Peirotén Á, Landete JM, Medina M (2012) Antimicrobial properties of probiotic strains isolated from breast-fed infants. J Funct Foods 4:542–551.  https://doi.org/10.1016/j.jff.2012.02.015 CrossRefGoogle Scholar
  22. 22.
    Schillinger U, Yousif NM, Sesar L, Franz CM (2003) Use of group-specific and RAPD-PCR analyses for rapid differentiation of Lactobacillus strains from probiotic yogurts. Curr Microbiol 47:453–456.  https://doi.org/10.1007/s00284-003-4067-8 CrossRefPubMedGoogle Scholar
  23. 23.
    Serafini F, Strati F, Ruas-Madiedo P, Turroni F, Foroni E, Duranti S, Milano F, Perotti A, Viappiani A, Guglielmetti S (2013) Evaluation of adhesion properties and antibacterial activities of the infant gut commensal Bifidobacterium bifidum PRL2010. Anaerobe 21:9–17.  https://doi.org/10.1016/j.anaerobe.2013.03.003 CrossRefPubMedGoogle Scholar
  24. 24.
    Shobharani P, Halami PM (2014) Cellular fatty acid profile and H+-ATPase activity to assess acid tolerance of Bacillus sp. for potential probiotic functional attributes. Appl Microbiol Biotechnol 98:9045–9058.  https://doi.org/10.1007/s00253-014-59813 CrossRefPubMedGoogle Scholar
  25. 25.
    Shetty SA, Marathe NP, Shouche YS (2013) Opportunities and challenges for gut microbiome studies in the Indian population. Microbiome.  https://doi.org/10.1186/2049-2618-1-24 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Sybesma W, Kort R, Lee Y-K (2015) Locally sourced probiotics, the next opportunity for developing countries? Trends Biotechnol 33(4):197–200CrossRefGoogle Scholar
  27. 27.
    Toscano M, De Vecchi E, Gabrieli A, Zuccotti GV, Drago L (2015) Probiotic characteristics and in vitro compatibility of a combination of Bifidobacterium breve M-16 V, Bifidobacterium longum subsp. infantis M-63 and Bifidobacterium longum subsp. longum BB536. Ann Microbiol 65:1079–1086.  https://doi.org/10.1007/s13213-014-0953-5 CrossRefGoogle Scholar
  28. 28.
    Turroni F, Foroni E, Motherway MOC, Bottacini F, Giubellini V, Zomer A, Ferrarini A, Delledonne M, Zhang Z, van Sinderen D (2010) Characterization of the serpin-encoding gene of Bifidobacterium breve 210B. Appl Environ Microbiol 76:3206–3219.  https://doi.org/10.1128/AEM.02938-09 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Turroni F, Serafini F, Foroni E, Duranti S, Motherway MOC, Taverniti V, Mangifesta M, Milani C, Viappiani A, Roversi T (2013) Role of sortase-dependent pili of Bifidobacterium bifidum PRL2010 in modulating bacterium–host interactions. Proc Natl Acad Sci USA 110:11151–11156.  https://doi.org/10.1073/pnas.1303897110 CrossRefPubMedGoogle Scholar
  30. 30.
    Ventura M, Turroni F, Motherway MOC, MacSharry J, van Sinderen D (2012) Host–microbe interactions that facilitate gut colonization by commensal bifidobacteria. Trends Microbiol 20:467–476.  https://doi.org/10.1016/j.tim.2012.07.002 CrossRefPubMedGoogle Scholar
  31. 31.
    Zinedine A, Faid M (2007) Isolation and characterization of strains of bifidobacteria with probiotic proprieties in vitro. World J Dairy Food Sci 2(1):28–34Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Academy of Scientific and Innovative Research (AcSIR)MysuruIndia
  2. 2.Microbiology and Fermentation Technology DepartmentCSIR-Central Food Technological Research InstituteMysuruIndia

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