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Updated: Mar 8, 2026

Individualized Reconstitution of Human Milk Microbiota: A Feasible Approach in Real-World Settings
Published on: February 7, 2025
High-temperature short-time pasteurisation of human breastmilk is efficient in retaining protein and reducing the
Daniel Klotz1,2, Mirjam Joellenbeck1,2, Karl Winkler3
1Division of Neonatology, Center for Pediatrics, Medical Center - University of Freiburg, Freiburg, Germany.
Aim:
Milk banks are advised to use Holder pasteurisation to inactivate the cytomegalovirus, but the process adversely affects the bioactive properties of human breastmilk. This study explored the antibacterial efficacy of an alternative high-temperature short-time (HTST) treatment of human breastmilk and its effect on marker proteins, compared with the Holder method.
Methods:
Breastmilk samples were obtained from 27 mothers with infants in a German neonatal intensive care unit. The samples were either heated to 62°C for five seconds using HTST or processed using Holder pasteurisation, at 63 ± 0.5°C for 30 minutes. Immunoglobulin A, lactoferrin, lysozyme, alkaline phosphatase and bile salt-stimulated lipase concentrations and bacterial colony-forming units/mL were measured before and after heating.
Results:
HTST-treated samples retained higher rates of immunoglobulin A (95% versus 83%), alkaline phosphatase (6% versus 0%) and bile salt-stimulated lipase (0.8% versus 0.4%) than Holder pasteurisation samples (all p < 0.01), but not lactoferrin (32% versus 20%, p = 0.18) and lysozyme (72% versus 65%, p = 1). No difference in antibacterial efficacy was noted between the two groups (p = 0.29).
Conclusion:
Using the HTST treatment protocol retained some of the bioactive properties of human breastmilk and appeared to have similar antibacterial efficacy to Holder pasteurisation.
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