Estimation of the probability of bacterial population survival: Development of a probability model to describe the variability in time to inactivation of Salmonella enterica


Kento Koyama, Hidekazu Hokunan, Mayumi Hasegawa, Shigenobu Koseki


Despite the development of numerous predictive microbial inactivation models, a model focusing on the variability in time to inactivation for a bacterial population has not been developed. Additionally, an appropriate estimation of the risk of there being any remaining bacterial survivors in foods after the application of an inactivation treatment has not yet been established. Here, Gamma distribution, as a representative probability distribution, was used to estimate the variability in time to inactivation for a bacterial population. Salmonella enterica serotype Typhimurium was evaluated for survival in a low relative humidity environment. We prepared bacterial cells with an initial concentration that was adjusted to 2 × 10n colony-forming units/2 μl (n = 1, 2, 3, 4, 5) by performing a serial 10-fold dilution, and then we placed 2 μl of the inocula into each well of 96-well microplates. The microplates were stored in a desiccated environment at 10–20% relative humidity at 5, 15, or 25 °C. The survival or death of bacterial cells for each well in the 96-well microplate was confirmed by adding tryptic soy broth as an enrichment culture. The changes in the death probability of the 96 replicated bacterial populations were described as a cumulative Gamma distribution. The variability in time to inactivation was described by transforming the cumulative Gamma distribution into a Gamma distribution. We further examined the bacterial inactivation on almond kernels and radish sprout seeds. Additionally, we described certainty levels of bacterial inactivation that ensure the death probability of a bacterial population at six decimal reduction levels, ranging from 90 to 99.9999%. Consequently, the probability model developed in the present study enables us to estimate the death probability of bacterial populations in a desiccated environment over time. This probability model may be useful for risk assessment to estimate the amount of remaining bacteria in a given sample.

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