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The viability of probiotics in water, breast milk, and infant formula


The aim of this study was to determine bacteriological stability of a probiotic mixture dispersed in various diluents. The commercially available probiotic (Infloran®), containing Bifidobacterium bifidum (109 CFU/250 mg tablet) and Lactobacillus acidophilus (109 CFU/250 mg tablet), was dispersed within expressed breast milk, sterile water, and infant formula and examined at temperatures of 4 and 21 °C. When stored at 4 °C, significant decreases (P < 0.05) in the level of L. acidophilus and B. bifidum were observed in expressed breast milk and sterile water after a 6-h period. However, when stored in infant formula, both strains remained stable over a 12-h period. When stored at 21 °C, a significant decrease (P < 0.05) was observed in the level of L. acidophilus in sterile water, expressed breast milk and infant formula throughout a 12-h period. However, no significant decrease was observed overtime in B. bifidum in all three diluents at this temperature.

Conclusion: Our findings suggest that, when stored at 4 °C, this probiotic product can remain at a stable condition for 6 h in sterile water and infant formula; however, the viability of the probiotic decreases significantly after this period of time. Administration of this probiotic in sterile water can be an acceptable alternative to dispersion and administration in expressed breast milk.

What is Known:

Administration of probiotics containing lactobacilli and bifidobacteria has become more widespread in neonatology, mainly as prophylaxis for the prevention of necrotising entercolitis in preterm infants.

Probiotic reconstitution, from its powder base, is not standardized and various diluents, including sterile water, breast milk, and infant formula, have been used.

What is New:

When stored at 4 °C, a probiotic containing lactobacilli and bifidobacteria remains at a stable microbological condition for up to 6 h in sterile water.

Administration of this probiotic dispersed in sterile water, followed by an EBM feed, can be an acceptable alternative to dispersion and administration in EBM.

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Fig. 1



Expressed breast milk


Lactobacillus selection


de Mann, Rogosa and Sharpe


Necrotising enter colitis


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The authors acknowledge the assistance of Carol-Anne O′ Shea and Maura Cahill from the Cork University Maternity Hospital (CUMH) for their help in providing the probiotic and for demonstrating probiotic administration in the NICU.


The authors are supported, in part, by the Science Foundation Ireland, through The APC Microbiome Institute (APC), and INFANTMET (Project (Ref No 10FDairy), funded by the Food Institutional Research Measure (FIRM) of the Department of Agriculture, Food and the Marine.

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Authors and Affiliations



Prof Tony Ryan, Eugene Dempsey, and Prof Paul W O’ Toole were responsible for substantial contributions to the conception and design of the work.

Claire Watkins acquired the data that was analyzed; Claire Watkins and Kiera Murphy analyzed and assisted with the interpretation of data of the work with Prof Paul Ross, Prof Catherine Stanton, and Prof Tony Ryan.

Drafting of the work and revising it critically for important intellectual content were carried out by Claire Watkins, Prof Tony Ryan, and Brendan P Murphy.

Agreement to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved by all authors.

Corresponding author

Correspondence to C. Anthony Ryan.

Ethics declarations

The research does not involve human or animals. This study did not require informed consent. Ethical approval for the study was granted from Clinical Research Ethics Committee of the Cork Teaching Hospitals. (ECM4(M)01/09/15& (EMC)(dddddddd)12/04/16.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by Patrick Van Reempts

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Watkins, C., Murphy, K., Dempsey, E.M. et al. The viability of probiotics in water, breast milk, and infant formula. Eur J Pediatr 177, 867–870 (2018).

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  • Probiotics
  • Expressed breast milk
  • Water
  • Preterm
  • Newborns