J Korean Med Sci.  2021 Mar;36(8):e58. 10.3346/jkms.2021.36.e58.

Comprehensive Analysis of the Effect of Probiotic Intake by the Mother on Human Breast Milk and Infant Fecal Microbiota

• Shin DY
• Park J
• Yi DY

• KMID: 2513533
• DOI: http://doi.org/10.3346/jkms.2021.36.e58

Abstract

Background
Human breast milk (HBM) contains optimal nutrients for infant growth. Probiotics are used to prevent disease and, when taken by the mother, they may affect infant microbiome as well as HBM. However, few studies have specifically investigated the effect of probiotic intake by the mother on HBM and infant microbiota at genus/species level. Therefore, we present a comprehensive analysis of paired HBM and infant feces (IF) microbiome samples before and after probiotic intake by HBM-producing mothers.
Methods
Lactating mothers were administered with Lactobacillus rhamnosus (n = 9) or Saccharomyces boulardii capsules (n = 9), for 2 months; or no probiotic (n = 7). Paired HBM and IF samples were collected before and after treatment and analyzed by next-generation sequencing.
Results
Forty-three HBM and 49 IF samples were collected and sequenced. Overall, in 43 HBM samples, 1,190 microbial species belonging to 684 genera, 245 families, 117 orders, and 56 classes were detected. In 49 IF samples, 372 microbial species belonging to 195 genera, 79 families, 42 orders, and 18 classes were identified. Eight of 20 most abundant genera in both HBM and IF samples overlapped: Streptococcus (14.42%), Lactobacillus, Staphylococcus, and Veillonella, which were highly abundant in the HBM samples; and Bifidobacterium (27.397%), Bacteroides, and Faecalibacterium, which were highly abundant in the IF samples. Several major bacterial genera and species were detected in the HBM and IF samples after probiotic treatment, illustrating complex changes in the microbiomes upon treatment.
Conclusion
This is the first Korean microbiome study in which the effect of different probiotic intake by the mother on the microbiota in HBM and IF samples was investigated. This study provides a cornerstone to further the understanding of the effect of probiotics on the mother and infant microbiomes.

Keyword

Human Breast Milk; Infant Feces; Breastfeeding; Probiotics; Microbiota; Korean

Figure

• 

  Fig. 1 Rarefaction analyses based on the 12 groups defined by sample type, probiotics, and treatment. X-axis is number of sequence reads per sample and Y-axis is observed OTUs (A) displays rarefraction analysis results of the six groups of human BM samples and (B) presents rarefraction analysis results of the six groups of infant fecal samples. X-axis indicates sequences per samples and Y-axis means observed OTUs.OTU = operational taxonomic unit, BM = breast milk.

• 

  Fig. 2 Distribution of microbial genera detected in HBM and IF samples. (A) Pie graph displaying the proportion of genera detected in HBM samples. (B) Pie graph displaying the proportion of genera detected in IF samples. (C) X-axis of graph shows genera name identified from both HBM and IF samples. Y-axis indicates relative proportion of each genus. Streptococcus (14.415%) max proportion identified from HBM samples, Bifidobacterium (27.397%) max proportion identified from IF samples, respectively.HBM = human breast milk, IF = infant fecal.

• 

  Fig. 3 Bacterial genera detected in the treatment groups, and their relative proportions in the microbiome. Readings before and after probiotic treatment are shown. (A) Relative genus proportions in HBM samples in the control group. (B) Relative genus proportions in IF samples in the control group. (C) Relative genus proportions in HBM samples in the L. rhamnosus group. (D) Relative genus proportions in IF samples in the L. rhamnosus group. (E) Relative genus proportions in HBM samples in the S. boulardii group. (F) Relative genus proportions in IF samples in the S. boulardii group. Y-axis indicates the proportion of genera of microbial species of which range is from 0 to 1.HBM = human breast milk, IF = infant fecal.

• 

  Fig. 4 Hierarchical clustering heat-map of genera that became more or less abundant under different experimental conditions. Microbial genera detected in the current study are shown along the y-axis. The experimental conditions are shown along the x-axis. Hierarchical clustering dendrograms are displayed on the left and top. Blue and red colors indicate a decrease or increase of a population in the microbiome, respectively, upon probiotic treatment.HBM = human breast milk, IF = infant fecal, Control = the control group, LR = the L. rhamnosus group, SB = the S. boulardii group.

• 

  Fig. 5 Comparison of microbial community structure under three experimental conditions. Blue boxes with transparent effect, analysis of HBM samples; green boxes, analysis of IF samples. Venn diagrams in each box show detected bacterial species before and after probiotic treatment. (A) Analysis of the control group samples. (B) Analysis of the L. rhamnosus group samples. (C) Analysis of the S. boulardii group samples.HBM = human breast milk, IF = infant fecal.

• 

  Fig. 6 Microbial species that were unique or common to the three experimental groups. Venn diagram in the center displays species detected under the three conditions. Species specific to the three groups and common species are listed.

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  Fig. 7 Proposed model for the establishment of infant fecal and human breast milk microbiomes. Blue boxes indicate factors that promote microbiome expansion and red boxes indicate environmental conditions that are detrimental to the microbiome. Grey arrow point to next generation sequencing microbiome analysis.

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