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Updated: Nov 26, 2025

Individualized Reconstitution of Human Milk Microbiota: A Feasible Approach in Real-World Settings
Published on: February 7, 2025
Metabolomic Profile of Personalized Donor Human Milk
Monica F Torrez Lamberti1, Evon DeBose-Scarlett1, Timothy Garret2
1Department of Microbiology and Cell Science, Genetics Institute, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32611, USA.
Abstract:
Human milk could be considered an active and complex mixture of beneficial bacteria and bioactive compounds. Since pasteurization drastically reduces the microbial content, we recently demonstrated that pasteurized donor human milk (DHM) could be inoculated with different percentages (10% and 30%) of mother's own milk (MOM) to restore the unique live microbiota, resulting in personalized milk (RM10 and RM30, respectively). Pasteurization affects not only the survival of the microbiota but also the concentration of proteins and metabolites, in this study, we performed a comparative metabolomic analysis of the RM10, RM30, MOM and DHM samples to evaluate the impact of microbial restoration on metabolite profiles, where metabolite profiles clustered into four well-defined groups. Comparative analyses of DHM and MOM metabolomes determined that over one thousand features were significantly different. In addition, significant changes in the metabolite concentrations were observed in MOM and RM30 samples after four hours of incubation, while the concentration of metabolites in DHM remained constant, indicating that these changes are related to the microbial expansion. In summary, our analyses indicate that the metabolite profiles of DHM are significantly different from that of MOM, and the profile of MOM may be partially restored in DHM through microbial expansion.
Insights
Restoring live microbiota in pasteurized donor human milk (DHM) with mother's own milk (MOM) partially restores its unique metabolite profiles. Microbial expansion drives these metabolic changes in personalized milk.
Area of Science:
- Human milk research
- Microbiology
- Metabolomics
Background:
- Human milk contains beneficial bacteria and bioactive compounds.
- Pasteurization reduces microbial content and alters metabolite profiles.
- Restoring microbiota in pasteurized donor milk (DHM) with mother's own milk (MOM) creates personalized milk (RM10, RM30).
Purpose of the Study:
- To compare the metabolomic profiles of personalized milk (RM10, RM30), mother's own milk (MOM), and pasteurized donor milk (DHM).
- To evaluate the impact of microbial restoration on metabolite profiles in DHM.
Main Methods:
- Comparative metabolomic analysis of DHM, MOM, RM10, and RM30 samples.
- Incubation of samples to observe metabolite concentration changes over time.
- Clustering of metabolite profiles to identify distinct groups.
Main Results:
- Metabolite profiles clustered into four distinct groups.
- Over one thousand metabolite features differed significantly between DHM and MOM.
- MOM and RM30 showed significant metabolite changes after incubation, unlike DHM, indicating microbial activity.
Conclusions:
- Pasteurized donor milk (DHM) has significantly different metabolite profiles compared to mother's own milk (MOM).
- Microbial expansion in personalized milk (RM) can partially restore the metabolite profile of MOM.
- Microbiota plays a crucial role in shaping the metabolome of human milk.

