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. 2025 Mar 25;14(4):332.
doi: 10.3390/biology14040332.

Human Milk Microbiome from Polish Women Giving Birth via Vaginal Delivery-Pilot Study

Agnieszka Chrustek  1 , Agnieszka Dombrowska-Pali  2 , Dorota Olszewska-Słonina  1 , Natalia Wiktorczyk-Kapischke  3 , Maciej W Socha  2 , Anna Budzyńska  3 , Iwona Sadowska-Krawczenko  4
Affiliations

Human Milk Microbiome from Polish Women Giving Birth via Vaginal Delivery-Pilot Study

Agnieszka Chrustek et al. Biology (Basel). .

Abstract

The human milk (HM) microbiome is variable and depends on maternal, perinatal, and cultural-environmental factors. The diversity of the HM microbiome is crucial in the development of the child. The aim of the study was to assess the prevalence of bacteria (using culture-based methods) of Polish women with normal BMI, giving birth on time through vaginal delivery.

Methods: The research material consisted of human milk and swabs from the areola and nipple, before and after breastfeeding, derived from Polish women (n = 86). Classic culture methods were used to obtain multiple bacteria. Species identification of the grown colonies was performed using MALDI TOF MS (Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry).

Results: 120 species of bacteria were isolated, mainly from the genus Streptococcus and Staphylococcus. Species specific only to human milk were identified (belonging to the following genera: Microbacterium, Shewanella, Psychrobacter, Aeromonas, Serratia, Buttiauxella, Lactobacillus, Bifidobacterium) as well as species specific only to areola and nipple swabs after breastfeeding (Acinetobacter lactucae, Moraxella catarrhalis, Corynebacterium pseudodiphtheriticum, Corynebacterium propinquim). It was confirmed that most species were present in all tested materials collected from one patient.

Conclusions: The analysis carried out showed the presence of bacteria in the human milk of Polish women, including strains of lactic acid bacteria. The human milk microbiota may significantly influence the formation of the infant's intestinal microbiota, including some key genera, i.e., Lactobacillus, Bifidobacterium, and Limosilactobacillus, which were also isolated from the tested samples. The data presented here provide new data on culturable bacterial species isolated from breast milk from Polish women giving birth via vaginal delivery and potential routes of transmission from the neonate's oral cavity.

Keywords: breastfeeding; culture-based method; human milk; microbiome; vaginal delivery.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Hypothetical sources of the human milk microbiome.
Figure 2
Figure 2
Percentage of bacterial genera isolated from the areola before breastfeeding. Other: Lacticaseibacillus, Schaalia, Leclercia, Cytobacillus, Lacticaseibacillus, Bacillus, Moraxella, and Escherichia.
Figure 3
Figure 3
Percentage of bacterial genera isolated from human milk. Other: Lacticaseibacillus, Bifidobacterium, Empeadobacter, Cytobacillus, Lacticaseibacillus, Microbacterium, Psychrobacter, Shewanella, Stenotrophomonas, Raoultella, Aeromonas, Serratia, Buttiauxella, Schaalia, Leclercia, and Pantonea.
Figure 4
Figure 4
Percentage of bacterial genera isolated from the areola after breastfeeding. Other: Lacticaseibacillus, Pantonea, Cutibacterium, Moraxella, Klebsiella, and Enterobacter.
Figure 5
Figure 5
Comparison of isolated genera from: the areola before and after breastfeeding and from human milk. Other: Lacticaseibacillus, Pantonea, Cutibacterium, Moraxella, Klebsiella, Enterobacter, Schaalia, Leclercia, Cytobacillus, Bacillus, Moraxella, Escherichia, Bifidobacterium, Empeadobacter, Microbacterium, Psychrobacter, Shewanella, Raoultella, Aeromonas, Serratia, Buttiauxella, Schaalia, and Leclercia.
Figure 6
Figure 6
Venn diagram showing unique and shared bacterial strains between human milk and areola swab samples before and after breastfeeding.
Figure 7
Figure 7
Dendrogram with heatmap showing common taxa of human milk and areola swab. Colormap from green to red shows increasing absolute abundance. Green shows lowest abundance and red the highest.
Figure 8
Figure 8
(A) Heatmap showing Pearson correlations between bacterial genera in the human milk. The legend shows the correlation coefficient values, while the axes show the genera of bacteria. (B) Heatmap showing Pearson correlations between bacterial genera in the areola swab before breastfeeding. The legend shows the correlation coefficient values, while the axes show the genera of bacteria. (C) Heatmap showing Pearson correlations between bacterial genera in the areola swab after breastfeeding. The legend shows the correlation coefficient values, while the axes show the genera of bacteria.
Figure 8
Figure 8
(A) Heatmap showing Pearson correlations between bacterial genera in the human milk. The legend shows the correlation coefficient values, while the axes show the genera of bacteria. (B) Heatmap showing Pearson correlations between bacterial genera in the areola swab before breastfeeding. The legend shows the correlation coefficient values, while the axes show the genera of bacteria. (C) Heatmap showing Pearson correlations between bacterial genera in the areola swab after breastfeeding. The legend shows the correlation coefficient values, while the axes show the genera of bacteria.
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