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. 2015 Jan 2;14(1):491-502.
doi: 10.1021/pr500759e. Epub 2014 Oct 28.

Human milk glycomics and gut microbial genomics in infant feces show a correlation between human milk oligosaccharides and gut microbiota: a proof-of-concept study

Maria Lorna A De Leoz  1 , Karen M Kalanetra, Nicholas A Bokulich, John S Strum, Mark A Underwood, J Bruce German, David A Mills, Carlito B Lebrilla
Affiliations

Human milk glycomics and gut microbial genomics in infant feces show a correlation between human milk oligosaccharides and gut microbiota: a proof-of-concept study (V体育2025版)

Maria Lorna A De Leoz et al. J Proteome Res. .

Abstract

Human milk oligosaccharides (HMOs) play a key role in shaping and maintaining a healthy infant gut microbiota. This article demonstrates the potential of combining recent advances in glycomics and genomics to correlate abundances of fecal microbes and fecal HMOs. Serial fecal specimens from two healthy breast-fed infants were analyzed by bacterial DNA sequencing to characterize the microbiota and by mass spectrometry to determine abundances of specific HMOs that passed through the intestinal tract without being consumed by the luminal bacteria. In both infants, the fecal bacterial population shifted from non-HMO-consuming microbes to HMO-consuming bacteria during the first few weeks of life VSports手机版. An initial rise in fecal HMOs corresponded with bacterial populations composed primarily of non-HMO-consuming Enterobacteriaceae and Staphylococcaeae. This was followed by decreases in fecal HMOs as the proportion of HMO-consuming Bacteroidaceae and Bifidobacteriaceae increased. Analysis of HMO structures with isomer differentiation revealed that HMO consumption is highly structure-specific, with unique isomers being consumed and others passing through the gut unaltered. These results represent a proof-of-concept and are consistent with the highly selective, prebiotic effect of HMOs in shaping the gut microbiota in the first weeks of life. The analysis of selective fecal bacterial substrates as a measure of alterations in the gut microbiota may be a potential marker of dysbiosis. .

Keywords: HMOs; Term infants; human milk; mass spectrometry; microbiota; oligosaccharides; prebiotic. V体育安卓版.

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Figure 1
Figure 1
Changes in oligosaccharide intensities and bacterial population in the feces of Infant A. (A) Percent change in H/D ratios of HMOs in the feces of Infant A at weeks 0, 1, 2, and 13. Intensities were obtained using MALDI FT-ICR MS with n = 3. H/D ratios were normalized to week 0, set at 100%. Each bar represents an oligosaccharide nominal mass. (B) Corresponding fecal bacterial population of Infant A using 16S rDNA pyrosequencing.
Figure 2
Figure 2
Infant A microbial population changes over the first 13 weeks based on next-generation sequencing. (A) Illumina and (B) pyrosequencing profile of the V4 and the V1–V3 regions of the 16S rRNA gene, respectively. The letters preceding the taxon are taxonomy identifiers: p (phylum), c (class), o (order), f (family), and g (genus).
Figure 3
Figure 3
Infant A microbial population changes over the first 13 weeks based on qPCR and Bif-TRFLP analyses. qPCR of (A) Total bacteria, Bifidobacteria, and Bacteroidales and (B) species-specific Bifidobacteria populations; (C) Bif-TRFLP profile of the total bifidobacteria community.
Figure 4
Figure 4
Changes in oligosaccharide intensities and bacterial population in the feces of Infant B. (A) Percent change in H/D ratios of HMOs in the feces of Infant B at weeks 0, 1, 2, and 14. Intensities were obtained using MALDI FT-ICR MS in (+) ion mode with n = 3. H/D ratios were normalized to week 0, set at 100%. Each bar represents an oligosaccharide nominal mass. (B) Corresponding fecal bacterial population of Infant B using 16S rDNA pyrosequencing.
Figure 5
Figure 5
Infant B microbial population changes over the first 14 weeks of life based on next-generation sequencing. (A) Illumina and (B) pyrosequencing profile of the V4 region and V1–V3 regions of the16S rRNA gene, respectively.
Figure 6
Figure 6
InfantB microbial population changes over the first 14 weeks of life based on qPCR analysis. qPCR of total bacteria, Bifidobacteria, and Bacteroidales. * Bifidobacteria are below the limit of detection (1 × 104 16S rRNA genes per gram of stool).
Figure 7
Figure 7
H/D ratios of two isomeric groups of oligosaccharides in the fecal HMO profile of Infant A. H/D ratios were calculated using nano-HLPC chip/TOF MS data. (A) Four isomers of m/z 856 ([M + H]+, M = 855.3220, second bar from the left in each week in Figure 1A). (B) Three isomers of m/z 538 ([M + 2H]2+, z = 2, M = 1074.3963, fourth bar from the left in Figure 1A). M = monoisotopic (neutral) mass.
Figure 8
Figure 8
LC–MS extracted ion chromatograms of m/z 538 ([M + 2H]2+, z = 2, M = 1074.3963) in the fecal HMO profile of Infant A using nano-HLPC chip/TOF MS. Chromatograms at (A) week 0, (B) week 1, (C) week 2, and (D) week 13. M = monoisotopic (neutral) mass.
Figure 9
Figure 9
H/D ratios of four isomeric groups of oligosaccharides in the fecal HMO profile of Infant A. H/D ratios were calculated using nano-HLPC chip/TOF MS data. (A) Five isomers of m/z 611 ([M + 2H]2+, z = 2, M = 1220.4542, sixth bar from the left in each week in Figure 1A). (B) Three isomers of m/z 684 ([M + 2H]2+, z = 2, M = 1366.5121, seventh bar from the left in Figure 1A). (C) Four isomers of m/z 794 ([M + 2H]2+, z = 2, M = 1585.5864, tenth bar from the left in Figure 1A). (D) Five isomers of m/z 867 ([M + 2H]2+, z = 2, M = 1731.6443, 11th bar from the left in Figure 1A). M = monoisotopic (neutral) mass.
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