In Vitro Comparative Analysis of Probiotic and Functional Attributes of Indigenous Isolates of Bifidobacteria
- 48 Downloads
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.
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).
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.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the Institute.
Informed consent was obtained from parents of all infants included in the study.
- 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
- 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
- 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.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.FAO/WHO (2002) WHO working group report on drafting guidelines for the evaluation of probiotics in food. FAO/WHO, LondonGoogle Scholar
- 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.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
- 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
- 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
- 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
- 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.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.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
- 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