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Tolerability and safety of the intake of bovine milk oligosaccharides extracted from cheese whey in healthy human adults

  • Jennifer T. Smilowitz (a1) (a2), Danielle G. Lemay (a3), Karen M. Kalanetra (a1), Elizabeth L. Chin (a1), Angela M. Zivkovic (a4), Melissa A. Breck (a1), J. Bruce German (a1) (a2), David A. Mills (a1), Carolyn Slupsky (a4) and Daniela Barile (a1) (a2)...

Mechanistic research suggests a unique evolutionary relationship between complex milk oligosaccharides and cognate bifidobacteria enriched in breast-fed infants. Bovine milk oligosaccharides (BMO) were recently identified as structurally and functionally similar to human milk oligosaccharides. The present single-blind three-way crossover study is the first to determine the safety and tolerability of BMO consumption by healthy human participants (n 12) and its effects on faecal microbiota and microbial metabolism. Participants consumed each supplement (placebo-control; low- and high-BMO doses) for eleven consecutive days, followed by a 2-week washout period prior to initiating the next supplement arm. Low and high BMO doses were consumed as 25 and 35 % of each individual's daily fibre intake, respectively. Safety and tolerability were measured using standardised questionnaires on gut and stomach discomfort and stool consistency. Faecal extracts were profiled for bacterial populations by next-generation sequencing (NGS) and bifidobacteria presence was confirmed using quantitative PCR. Urine was analysed for changes in microbial metabolism using nuclear magnetic resonance spectroscopy (1H-NMR). Consumption of both the low and high BMO doses was well tolerated and did not change stool consistency from baseline. Multivariate analysis of the NGS results demonstrated no change in faecal microbiota phyla among the placebo-control and BMO supplement groups. In conclusion, BMO supplementation was well tolerated in healthy adults and has the potential to shift faecal microbiota toward beneficial strains as part of a synbiotic treatment with probiotic cultures that selectively metabolise oligosaccharides.

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This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (, which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.
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* Corresponding author: D. Barile, email
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1. Garrido, D, Kim, JH, German, JB, et al. (2011) Oligosaccharide binding proteins from Bifidobacterium longum subsp. infantis reveal a preference for host glycans. PLoS ONE 6, e17315.
2. Coppa, G, Pierani, P, Zampini, L, et al. (2001) Characterization of oligosaccharides in milk and feces of breast-fed infants by high-performance anion-exchange chromatography. In Bioactive Components of Human Milk, pp. 307314 [Newburg, DS, editor]. New York: Springer.
3. Chaturvedi, P, Warren, CD, Buescher, CR, et al. (2001) Survival of human milk oligosaccharides in the intestine of infants. In Bioactive Components of Human Milk, pp. 315323 [Newburg, DS, editor]. New York: Springer.
4. Engfer, MB, Stahl, B, Finke, B, et al. (2000) Human milk oligosaccharides are resistant to enzymatic hydrolysis in the upper gastrointestinal tract. Am J Clin Nutr 71, 15891596.
5. LoCascio, RG, Niñonuevo, MR, Kronewitter, SR, et al. (2009) A versatile and scalable strategy for glycoprofiling bifidobacterial consumption of human milk oligosaccharides. Microb Biotechnol 2, 333342.
6. Ward, RE, Ninonuevo, M, Mills, DA, et al. (2007) In vitro fermentability of human milk oligosaccharides by several strains of bifidobacteria. Mol Nutr Food Res 51, 13981405.
7. Sela, DA, Garrido, D, Lerno, L, et al. (2012) Bifidobacterium longum subsp. infantis ATCC 15697 α-fucosidases are active on fucosylated human milk oligosaccharides. Appl Environ Microbiol 78, 795803.
8. Sela, DA, Chapman, J, Adeuya, A, et al. (2008) The genome sequence of Bifidobacterium longum subsp. infantis reveals adaptations for milk utilization within the infant microbiome. Proc Natl Acad Sci U S A 105, 1896418969.
9. Tao, N, DePeters, EJ, German, JB, et al. (2009) Variations in bovine milk oligosaccharides during early and middle lactation stages analyzed by high-performance liquid chromatography-chip/mass spectrometry. J Dairy Sci 92, 29913001.
10. Barile, D, Marotta, M, Chu, C, et al. (2010) Neutral and acidic oligosaccharides in Holstein-Friesian colostrum during the first 3 days of lactation measured by high performance liquid chromatography on a microfluidic chip and time-of-flight mass spectrometry. J Dairy Sci 93, 39403949.
11. Nwosu, CC, Aldredge, DL, Lee, H, et al. (2012) Comparison of the human and bovine milk N-glycome via high-performance microfluidic chip liquid chromatography and tandem mass spectrometry. J Proteome Res 11, 29122924.
12. Tao, N, DePeters, EJ, Freeman, S, et al. (2008) Bovine milk glycome. J Dairy Sci 91, 37683778.
13. Barile, D, Tao, N, Lebrilla, CB, et al. (2009) Permeate from cheese whey ultrafiltration is a source of milk oligosaccharides. Int Dairy J 19, 524530.
14. LoCascio, RG, Ninonuevo, MR, Freeman, SL, et al. (2007) Glycoprofiling of bifidobacterial consumption of human milk oligosaccharides demonstrates strain specific, preferential consumption of small chain glycans secreted in early human lactation. J Agric Food Chem 55, 89148919.
15. Bouhnik, Y, Vahedi, K, Achour, L, et al. (1999) Short-chain fructo-oligosaccharide administration dose-dependently increases fecal bifidobacteria in healthy humans. J Nutr 129, 113116.
16. Baecke, J, Burema, J & Frijters, J (1982) A short questionnaire for the measurement of habitual physical activity in epidemiological studies. Am J Clin Nutr 36, 936942.
17. Institute of Medicine of the National Academies (2005) Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, DC: National Academies Press.
18. Pedersen, A, Sandström, B & Van Amelsvoort, JMM (1997) The effect of ingestion of inulin on blood lipids and gastrointestinal symptoms in healthy females. Br J Nutr 78, 215222.
19. Lewis, SJ & Heaton, KW (1997) Stool form scale as a useful guide to intestinal transit time. Scand J Gastroenterol 32, 920924.
20. Caporaso, JG, Lauber, CL, Walters, WA, et al. (2011) Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. Proc Nat Acad Sci U S A 108, 45164522.
21. Penders, J, Vink, C, Driessen, C, et al. (2005) Quantification of Bifidobacterium spp., Escherichia coli and Clostridium difficile in faecal samples of breast-fed and formula-fed infants by real-time PCR. FEMS Microbiol Lett 243, 141147.
22. Caporaso, JG, Kuczynski, J, Stombaugh, J, et al. (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7, 335336.
23. Edgar, RC (2010) Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26, 24602461.
24. Wang, Q, Garrity, GM, Tiedje, JM, et al. (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73, 52615267.
25. Cole, JR, Wang, Q, Cardenas, E, et al. (2009) The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res 37, D141D145.
26. DeSantis, TZ, Hugenholtz, P, Larsen, N, et al. (2006) Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol 72, 50695072.
27. Caporaso, JG, Bittinger, K, Bushman, FD, et al. (2010) PyNAST: a flexible tool for aligning sequences to a template alignment. Bioinformatics 26, 266267.
28. Slupsky, CM, Rankin, KN, Fu, H, et al. (2009) Pneumococcal pneumonia: potential for diagnosis through a urinary metabolic profile. J Proteome Res 8, 55505558.
29. Slupsky, CM, Rankin, KN, Wagner, J, et al. (2007) Investigations of the effects of gender, diurnal variation, and age in human urinary metabolomic profiles. Anal Chem 79, 69957004.
30. Weljie, AM, Newton, J, Mercier, P, et al. (2006) Targeted profiling: quantitative analysis of 1H NMR metabolomics data. Anal Chem 78, 44304442.
31. Smilowitz, JT, O'Sullivan, A, Barile, D, et al. (2013) The human milk metabolome reveals diverse oligosaccharide profiles. J Nutr 143, 17091718.
32. Segata, N, Izard, J, Waldron, L, et al. (2011) Metagenomic biomarker discovery and explanation. Genome Biol 12, R60.
33. Kruse, HP, Kleessen, B & Blaut, M (1999) Effects of inulin on faecal bifidobacteria in human subjects. Br J Nutr 82, 375382.
34. Kolida, S, Meyer, D & Gibson, GR (2007) A double-blind placebo-controlled study to establish the bifidogenic dose of inulin in healthy humans. Eur J Clin Nutr 61, 11891195.
35. Bouhnik, Y, Achour, L, Paineau, D, et al. (2007) Four-week short chain fructo-oligosaccharides ingestion leads to increasing fecal bifidobacteria and cholesterol excretion in healthy elderly volunteers. Nutr J 6, 42.
36. Rao, VA (2001) The prebiotic properties of oligofructose at low intake levels. Nutr Res 21, 843848.
37. Tuohy, KM, Finlay, RK, Wynne, AG, et al. (2001) A human volunteer study on the prebiotic effects of HP-inulin – faecal bacteria enumerated using fluorescent in situ hybridisation (FISH). Anaerobe 7, 113118.
38. Meli, F, Puccio, G, Cajozzo, C, et al. (2014) Growth and safety evaluation of infant formulae containing oligosaccharides derived from bovine milk: a randomized, double-blind, noninferiority trial. BMC Pediatr 14, 306.
39. Vázquez-Fresno, R, Llorach, R, Urpi-Sarda, M, et al. (2014) Metabolomic pattern analysis after Mediterranean diet intervention in a nondiabetic population: a 1- and 3-year follow-up in the PREDIMED study. J Proteome Res 14, 531540.
40. Higgins, J, Ramsay, C, Reeves, BC, et al. (2013) Issues relating to study design and risk of bias when including non-randomized studies in systematic reviews on the effects of interventions. Res Synth Methods 4, 1225.
41. Faith, JJ, Guruge, JL, Charbonneau, M, et al. (2013) The long-term stability of the human gut microbiota. Science 341, 1237439.
42. McNulty, NP, Yatsunenko, T, Hsiao, A, et al. (2011) The impact of a consortium of fermented milk strains on the gut microbiome of gnotobiotic mice and monozygotic twins. Sci Transl Med 3, 106ra106106ra106.
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Journal of Nutritional Science
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