Related Experiment Video
Updated: Jul 19, 2026

Individualized Reconstitution of Human Milk Microbiota: A Feasible Approach in Real-World Settings
Published on: February 7, 2025
Bacterial safety of flash-heated and unheated expressed breastmilk during storage
K Israel-Ballard1, A Coutsoudis, C J Chantry
1Division of Epidemiology, School of Public Health, University of California, Berkeley, CA 94720-7360, USA. ballardk@berkeley.edu
Heat-treated breastmilk is one infant-feeding option recommended by the WHO to reduce mother-to-child transmission of HIV in developing countries. Flash-heat, a simple pasteurization method that a mother could perform in her home, has been shown to inactivate cell-free HIV-1. Since heating may affect the naturally occurring antimicrobial properties found in breastmilk, storing heated breastmilk may present a safety issue in resource-poor settings due to lack of refrigeration and potential contamination. To address this, we investigated the ability of flash-heat to eliminate bacteria and to prevent growth over time compared with unheated breastmilk. We collected breastmilk samples from 38 HIV positive mothers in South Africa and aliquoted them to flash-heated and unheated controls. Samples were stored at room temperature for 0, 2, 6 and 8 h and then plated and incubated for 24 h at 37 degrees C in CO(2). We performed total colony counts and identified Escherichia coli, Staphylocuccus aureus and Group A and Group B streptococci. Unheated samples had a significantly higher number of samples positive for bacterial growth at each time point (p < 0.0001), as well as mean colony-forming units (CFU)/ml in those samples that were positive at each time point (p < 0.0001). In addition, unheated samples had a significantly higher rate of bacterial propagation over time than flash-heated samples when comparing log values of CFU/ml across 0-8 h (p < 0.005). No pathogenic growth was observed in the flash-heated samples, while the unheated samples showed growth of E. coli (n = 1) and S. aureus (n = 6). Our data suggest that storage of flash-heated breastmilk is safe at room temperature for up to 8 h.
Heat-treated breastmilk is one infant-feeding option recommended by the WHO to reduce mother-to-child transmission of HIV in developing countries. Flash-heat, a simple pasteurization method that a mother could perform in her home, has been shown to inactivate cell-free HIV-1. Since heating may affect the naturally occurring antimicrobial properties found in breastmilk, storing heated breastmilk may present a safety issue in resource-poor settings due to lack of refrigeration and potential contamination. To address this, we investigated the ability of flash-heat to eliminate bacteria and to prevent growth over time compared with unheated breastmilk. We collected breastmilk samples from 38 HIV positive mothers in South Africa and aliquoted them to flash-heated and unheated controls. Samples were stored at room temperature for 0, 2, 6 and 8 h and then plated and incubated for 24 h at 37 degrees C in CO(2). We performed total colony counts and identified Escherichia coli, Staphylocuccus aureus and Group A and Group B streptococci. Unheated samples had a significantly higher number of samples positive for bacterial growth at each time point (p < 0.0001), as well as mean colony-forming units (CFU)/ml in those samples that were positive at each time point (p < 0.0001). In addition, unheated samples had a significantly higher rate of bacterial propagation over time than flash-heated samples when comparing log values of CFU/ml across 0-8 h (p < 0.005). No pathogenic growth was observed in the flash-heated samples, while the unheated samples showed growth of E. coli (n = 1) and S. aureus (n = 6). Our data suggest that storage of flash-heated breastmilk is safe at room temperature for up to 8 h.
Related Concept Videos
Pasteurization and Food Preservation
Physical Methods for Controlling Microbial Growth: Temperature
Methods of Sterilization I: Physical Methods
Steam sterilization uses non-toxic, low-cost moist heat in the form of saturated steam under pressure, which is fast, microbicidal, and sporicidal, and quickly warms and penetrates fabrics. Autoclaves, or steam sterilizers, expose each item to direct steam contact for a predetermined time at the necessary...
Key Techniques in Microbiology

