Evaluation of Potential Probiotics Isolated from Human Milk and Colostrum

Abstract

Several studies have demonstrated a diversity of bacterial species in human milk, even in aseptically collected samples. The present study evaluated potential probiotic bacteria isolated from human milk and associated maternal variables. Milk samples were collected from 47 healthy women and cultured on selective and universal agar media under aerobic and anaerobic conditions. Bacterial isolates were counted and identified by Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight mass spectrometry and then tested for probiotic properties. Total bacteria in human milk ranged from 1.5 to 4.0 log10 CFU/mL. The higher bacterial counts were found in colostrum (mean = 3.9 log10 CFU/mL, 95% CI 3.14–4.22, p = 0.00001). The most abundant species was Staphylococcus epidermidis (n = 76). The potential probiotic candidates were Lactobacillus gasseri (n = 4), Bifidobacterium breve (n = 1), and Streptococcus salivarius (n = 4). Despite the small sample size, L. gasseri was isolated only in breast milk from mothers classified into a normal weight range and after a vaginally delivered partum. No potential probiotics showed antagonism against pathogens, but all of them agglutinated different pathogens. Nine bacterial isolates belonging to the species L. gasseri, B. breve, and S. salivarius were selected as potential probiotics. The present study confirms the presence in breast milk of a bacterial microbiota that could be the source of potential probiotic candidates to be used in the formula of simulated maternal milk.

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

Fig. 1

References

  1. 1.

    World Health Organization, Unicef. 2009. Baby-friendly hospital initiative: revised, updated and expanded for integrated care. Geneva, Switzerland: Author.

  2. 2.

    Jeurink PV, Bergenhenegouwen JV, Jiménez E, Knippels LMJ, Fernández L, Garssen J, Knol J, Rodríguez JM, Martin R (2013) Human milk: a source of more life than we imagine. Benef Microbes 4:7–30

    Article  Google Scholar 

  3. 3.

    McGuire MK, McGuire MA (2015) Human milk: mother nature’s prototypical probiotic food? Adv Nutr 6:112–123

    CAS  Article  Google Scholar 

  4. 4.

    Coppa GV, Bruni S, Morelli L, Soldi S, Gabrielli O (2004) The first prebiotics in humans. Human milk oligosaccharides J Clin Gastroenterol 38:S80–S83

    CAS  Article  Google Scholar 

  5. 5.

    Hunt KM, Foster JA, Forney LJ, Schütte UME, Beck DL, Abdo Z, Fox LK, Willians JE, McGuire MK, McGuire MA (2011) Characterization of the diversity and temporal stability of bacterial communities in human milk. PLoS One 6:e21313

    CAS  Article  Google Scholar 

  6. 6.

    Jiménez E, Andrés J, Manrique M, Pareja-Tobes P, Tobes R, Martínez-Blanch JF, Codoñer FM, Ramón D, Fernández L, Rodríguez JM (2015) Metagenomic analysis of milk of healthy and mastitis-suffering women. J Hum Lact 3:4016–4015

    Google Scholar 

  7. 7.

    Orban JI, Patterson JA (2000) Modification of the phosphoketolase assays for rapid identification of bifidobacteria. J Microbiol Methods 40:221–224

    CAS  Article  Google Scholar 

  8. 8.

    TeKippe EM, Burnham CAD (2014) Evaluation of the Bruker Biotyper and VITEK MS MALDI-TOF MS systems for the identification of unusual and/or difficult-to-identify microorganisms isolated from clinical specimens. Eur J Clin Microbiol Infect Dis 33:2163–2171

    Article  Google Scholar 

  9. 9.

    Lévesque S, Dufresne PJ, Soualhine H, Domingo MC, Bekal S, Lefebvre B, Tremblay C (2015) A side by side comparison of Bruker Biotyper and VITEK MS: utility of MALDI-TOF MS technology for microorganism identification in a public health reference laboratory. PLoS One 10(12):e0144878

    Article  Google Scholar 

  10. 10.

    Pérez-Sotelo LS, Talavera-Rojas M, Monroy-Salazar HG, Lagunas-Bernabé S, Cuarón-Ibargüengoytia JA, Jimenez RM, Vázquez-Chagoyán JC (2005) In vitro evaluation of the binding capacity of Saccharomyces cerevisiae Sc47 to adhere to the wall of Salmonella spp. Rev Latinamer Microbiol 47:70–75

    Google Scholar 

  11. 11.

    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

    CAS  Article  Google Scholar 

  12. 12.

    Noriega L, Gueimonde M, Sánchez B, Margolles A, Reyes-Gavilán CG (2004) Effect of the adaptation to high bile salts concentrations on glycosidic activity, survival at low PH and cross-resistance to bile salts in Bifidobacterium. Int J Food Microbiol 94:79–86

    CAS  Article  Google Scholar 

  13. 13.

    Farias FF, Lima FL, Carvalho MAR, Nicoli JR, Farias LM (2001) Influence of isolation site, laboratory handling and growth stage on oxygen tolerance of Fusobacterium strains. Anaerobe 7:271–276

    Article  Google Scholar 

  14. 14.

    Solís G, Reyes-Gavilan CG, Fernández N, Margolles A, Gueimonde M (2010) Establishment and development of lactic acid bacteria and bifidobacteria microbiota in breast milk and the infant gut. Anaerobe 16:307–310

    Article  Google Scholar 

  15. 15.

    Patras KA, Wescombe PA, Rösler B, Hale JD, Tagg JR, Doran KS (2015) Streptococcus salivarius K12 limits group B streptococcus vaginal colonization. Infect Immun 83:3438–3444

    CAS  Article  Google Scholar 

  16. 16.

    Thomas DW, Greer FR (2010) Committee on nutrition; section on gastroenterology, hepatology and nutrition probiotics and prebiotics in pediatrics. Pediatrics 126:1217–1231

    Article  Google Scholar 

  17. 17.

    Li Y, Shimizu T, Hosaka A, Kaneko N, Ohtsuka Y, Yamashiro Y (2004) Effects of Bifidobacterium breve supplementation on intestinal flora of low birth weight infants. Pediat Int 46:509–515

    Article  Google Scholar 

  18. 18.

    Jiménez E, Delgado S, Fernández L, García N, Albújar M, Gómez A, Rodríguez JM (2008) Assessment of the bacterial diversity of human colostrum and screening of staphylococcal and enterococcal populations for potential virulence factors. Res Microbiol 159:595–601

    Article  Google Scholar 

  19. 19.

    Obermajer T, Lipoglavšek L, Tompa G, Treven P, Lorbeg PM, Matijašić BB, Rogelj I (2015) Colostrum of healthy Slovenian mothers: microbiota composition and bacteriocin gene prevalence. PLoS One 10:e0123324

    Article  Google Scholar 

  20. 20.

    Picciano MF (1998) Human milk: nutritional aspects of a dynamic food. Biol Neonate 74:84–93

    CAS  Article  Google Scholar 

  21. 21.

    Kumar H, Toit E, Kulkarni A, Aakko J, Linderborg KM, Zhang Y, Nicol MP, Isolauri E, Yang B, Collado MC, Salminen S (2016) Distinct patterns in human milk microbiota and fatty acid profiles across specific geographic locations. Front Microbiol 7. doi:10.3389/fmicb.2016.01619

  22. 22.

    Burton JP, Cowley S, Simon RR, McKinney J, Wescombe PA, Tagg JR (2011) Evaluation of safety and human tolerance of the oral probiotic Streptococcus salivarius K12: a randomized, placebo-controlled, double-blind study. Food Chem Toxicol 49:2356–2364

    CAS  Article  Google Scholar 

  23. 23.

    Million M, Angelakis E, Paul M, Armougoma F, Leibovici L, Raoult D (2012) Comparative meta-analysis of the effect of Lactobacillus species on weight gain in humans and animals. Microb Pathog 53:100–108

    Article  Google Scholar 

  24. 24.

    Ley RE, Turnbaugh PJ, Klein S, Gordon JI (2006) Human gut microbes associated with obesity. Nat Microbiol 444:1022–1023

    CAS  Google Scholar 

  25. 25.

    Collado MC, Laitinen K, Salminen S, Isolauri E (2016) Maternal weight and excessive weight gain during pregnancy modify the immunomodulatory potential of breast milk. Clin Investig 72:77–85

    Google Scholar 

  26. 26.

    Barretto C, Alvarez-Martin P, Foata F, Renault P, Berger B (2012) Genome sequence of the lantibiotic bacteriocin producer Streptococcus salivarius strain K12. J Bacteriol 194:5959–5960

    CAS  Article  Google Scholar 

  27. 27.

    Martín V, Maldonado-Barragán A, Moles L, Rodriguez-Baños M, Del Campo R, Fernández L, Rodríguez JM, Jiménez E (2012) Sharing of bacterial strains between breast milk and infant feces. J Hum Lact 28:36–44

    Article  Google Scholar 

  28. 28.

    Kozaka K, Charbonneaua D, Sanozky-Dawesb R, Klaenhammerb T (2015) Characterization of bacterial isolates from the microbiota of mothersʼ breast milk and their infants. Gut Microbiol 6:341–351

    Article  Google Scholar 

  29. 29.

    Tiago FCP, Martins FS, Souza ELS, Pimenta PFP, Araújo HRC, Castro IM, Brandão RL, Nicoli JR (2012) Adhesion to the yeast cell surface as a mechanism for trapping pathogenic bacteria by Saccharomyces probiotics. J Med Microbiol 61:1194–1207

    CAS  Article  Google Scholar 

  30. 30.

    Selle K, Klaenhammer TR (2013) Genomic and phenotypic evidence for probiotic influences of Lactobacillus gasseri on human health. FEMS Microbiol Rev 37:915–935

    CAS  Article  Google Scholar 

  31. 31.

    Mogna L, Del Piano M, Mogna G (2014) Capability of the two microorganisms Bifidobacterium breve b632 and Bifidobacterium breve br03 to colonize the intestinal microbiota of children. J Clin Gastroenterol 48:S37–S39

    Article  Google Scholar 

  32. 32.

    Martín R, Olivares M, Marín ML, Fernández L, Xaus J, Rodríguez JM (2005) Probiotic potential of 3 lactobacilli strains isolated from breast milk. J Hum Lact 21:8–17

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from Brazilian National Council for Scientific and Technological Development (CNPq), Ministry of Science and Technology, and Foundation for Research Support of the State of Minas Gerais (FAPEMIG), Brazil. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript. QSD was the recipient of a fellowship from CNPq.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Flaviano S. Martins.

Ethics declarations

The volunteers were recruited from a public hospital in Belo Horizonte, Brazil, and a written informed consent was obtained from each individual. All procedures were performed after approval by the Ethical Committee (CAAE-31473714.0.0000.5149).

Conflict of Interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Damaceno, Q.S., Souza, J.P., Nicoli, J.R. et al. Evaluation of Potential Probiotics Isolated from Human Milk and Colostrum. Probiotics & Antimicro. Prot. 9, 371–379 (2017). https://doi.org/10.1007/s12602-017-9270-1

Download citation

Keywords

  • Human milk
  • Colostrum
  • Partum
  • Body mass index
  • Probiotics
  • L. gasseri