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Updated: May 13, 2026

Formulations for Freeze-drying of Bacteria and Their Influence on Cell Survival
Published on: August 3, 2013
Long-Term Survival of Cronobacter sakazakii, Salmonella enterica, Listeria monocytogenes, and Enterobacter spp. in
Megan L Fay1, Bashayer A Khouja1, Robert Newkirk1
1Division of Food Processing Science and Technology, U. S. Food and Drug Administration, Bedford Park, Illinois, USA.
Abstract:
Powdered infant formula (PIF) has previously been shown to support the survival of bacterial foodborne pathogens including Cronobacter sakazakii, Listeria monocytogenes, and Salmonella enterica at various storage temperatures. However, the effect of long-term storage on bacterial survival under different relative humidity (RH) conditions is not well understood. In this study, milk-based powdered infant formula (water activity (aw) = 0.27) was inoculated individually with cocktails of Salmonella enterica, Enterobacter spp., Cronobacter sakazakii, or Listeria monocytogenes and stored at 25 °C under controlled relative humidity (RH) levels of 23, 33, 43, 53, or 75%. Bacterial populations and PIF aw were measured at intervals up to 360 days. The highest aw of 0.39 was recorded for PIF stored under 75% RH after 90 d or at 53% RH after 360 d; however, these levels also resulted in significant bacterial population reductions of >1 log CFU/g after 60-120 d and 60-180 d, respectively, with L. monocytogenes being the least desiccation-resistant of those tested. RH levels between 23 and 43% did not result in a significant change in aw over 360 d; however, bacterial populations did decrease, but to a lesser extent than for the higher RH levels. Population reductions for all bacteria were ≤3.50 log CFU/g after 360 d. All four bacteria examined in this study persisted in PIF for ≥90 d without a significant decrease in population regardless of RH storage conditions.
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